Rifamycin analogs and uses thereof

ABSTRACT

The present invention features rifamycin analogs that can be used as therapeutics for treating or preventing a variety of microbial infections. In one form, the analogs are acetylated at the 25-position, as is rifamycin. In another form, the analogs are deacetylated at the 25-position. In yet other forms, benzoxazinorifamycin, benzthiazinorifamycin, and benzdiazinorifamycin analogs are derivatized at various positions of the benzene ring, including 3′-hydroxy analogs, 4′-and/or 6′ halo and/or alkoxy analogs, and various 5′ substituents that incorporate a cyclic amine moiety.

CROSS REFERENCE TO RELATED APPLICATIONS

Applicants claim the priority benefit under 35 U.S.C. 119(e) of priorprovisional application serial no. 60/497,058 filed Aug. 22, 2003, thedisclosure of which is incorporated herein by reference.

BACKGROUND OF THE INVENTION

The present invention relates to the field of antimicrobial agents.

The use of antibiotics by humans can be seen as an evolutionaryexperiment of enormous magnitude, a window from which to viewnot-quite-natural selection operating in real time. Within 50 years, thenumber of species and strains of pathogenic and commensal bacteriaresistant to antibiotics and the number of antibiotics to which they areresistant has increased virtually monotonically world-wide. As a result,infections that had been readily treatable by chemotherapy may no longerbe so. It is clear that the evolution and spread of resistance can beattributed to the use and overuse of antibiotics. Increased resistanceof bacterial infections to antibiotic treatment has been extensivelydocumented and has now become a generally recognized medical problem,particularly with nosocomial infections. See, for example, Jones et al.,Diagn. Microbiol. Infect. Dis. 31:379-388, 1998; Murray, Adv. Intern.Med. 42:339-367, 1997; and Nakae, Microbiologia 13:273-284, 1997.

Throughout the developed world there is public and governmental concernabout the increasing prevalence of antimicrobial resistance tochemotherapy in bacteria that cause diseases in humans. Many pathogensexist for which there are few effective treatments, and the number ofstrains resistant to available drugs is continually increasing. Newantimicrobial agents and improved methods are thus needed for thetreatment and prevention of infections by such pathogens.

SUMMARY OF THE INVENTION

The present invention features rifamycin analogs that can be used astherapeutics for treating or preventing a variety of microbialinfections.

Accordingly in a first aspect, the invention features a compound offormula (I):

In formula (I), A is H, OH, O—(C₁-C₆ alkyl), or O—(C₁-C₄ alkaryl); W isO, S, or NR¹, where R¹ is H or C₁-C₆ alkyl; X is H or COR², where R² isC₁-C₆ alkyl, which can be substituted with from 1 to 5 hydroxyl groups,or O—(C₃-C₇ alkyl, can be substituted with from 1 to 4 hydroxyl groups;each of Y and Z is independently H, C₁-C₆ alkoxy, or Hal; and R⁴ has thefollowing formula:

For the formula that represents R⁴, when each of m and n is 1, each ofR⁵ and R⁶ is H, or R⁵ and R⁶ together are ═O; R⁷ and R¹⁰ together form asingle bond or a C₁-C₃ linkage, R⁷ and R¹² together form a single bondor a C₁-C₂ linkage, or R⁷ and R¹⁴ together form a single bond or a C₁linkage; R⁸ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R⁸ and R¹² togetherform a single bond, or R⁸ and R⁹ together are ═N—OR⁸, where R¹⁸ is H,C₁-C₆ alkyl, or C₁-C₄ alkaryl; R⁹ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl,or R⁹ and R⁸ together are ═N—OR¹⁸; R¹⁰ is H, C₁-C₆ alkyl, or C₁-C₄alkaryl, or R¹⁰ and R¹⁷ together form a C₁-C₃ alkyl linkage, or R¹⁰ andR¹¹ together are ═O; R¹¹ is H, R¹² is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl;each of R¹³ and R¹⁵ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl; R¹⁴ is H, C₁-C₆alkyl, or C₁-C₄ alkaryl; R¹⁶ is H, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₆-C₁₂aryl, heteroaryl, C₁-C₄ alkaryl, or C₁-C₄ alkheteroaryl, or R¹⁶ and R¹²together form a C₂-C₄ alkyl linkage, or R¹⁶ and R¹⁰ together form aC₁-C₂ alkyl linkage; and R¹⁷ is H, C₁-C₆ alkyl, COR¹⁹, CO₂R¹⁹, orCONHR¹⁹, CSR¹⁹, COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or SO₂NHR¹⁹, where R¹⁹is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄alkheteroaryl, and where each alkyl linkage of 2 carbons or more maycontain a non-vicinal O, S, or N(R²³) where R²³ is H, C₁-C₆ alkyl,COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R₂₄, orSO₂NHR²⁴, where R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl,heteroaryl, or C₁-C₄ alkheteroaryl.

When m is 0 and n is 1, R⁷ and R¹⁰ together form a single bond or aC₁-C₄ linkage, R⁷ and R¹² together form a single bond or a C₁-C₃linkage, or R⁷ and R¹⁴ together form a single bond or a C₁-C₂ linkage;each of R⁸, R⁹, and R¹¹ is H; R¹⁵ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl;R¹⁰ is H; R¹² is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, R¹² and R¹³ togetherform a —CH₂CH₂—linkage, or R¹² and R¹⁶ together form a C₂-C₄ alkyllinkage; R¹³ is H, C₁-C₆ alkyl, C₁-C₄ alkaryl; R¹⁴ is H, C₁-C₆ alkyl, orC₁-C₄ alkaryl; R¹⁶ is H, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₆-C₁₂ aryl,heteroaryl, C₁-C₄ alkaryl, or C₁-C₄ alkheteroaryl, or R¹⁶ and R¹²together form a C₂-C₄ alkyl linkage; and R¹⁷is H, C₁-C₆ alkyl, COR¹⁹,CO₂R¹⁹, or CONHR¹⁹, CSR¹⁹, COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or SO₂NHR¹⁹,where R¹⁹ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, orC₁-C₄ alkheteroaryl, and where each alkyl linkage of 2 carbons or moremay contain a non-vicinal O, S, or N(R²³) where R²³ is H, C₁-C₆ alkyl,COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, orSO₂NHR²⁴, where R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl,heteroaryl, or C₁-C₄ alkheteroaryl.

Alternatively, for a compound of formula (I), A is OH; X is H; W, Y, andZ are as described above; and R⁴ is selected from the following groups:

where R²¹ is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl, orC₁-C₄ alkheteroaryl, R²⁰ is H, C₁-C₆ alkyl, COR¹⁹, CO₂R¹⁹, or CONHR¹⁹,CSR¹⁹, COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or SO₂NHR ₁₉, where R¹⁹ is C₁-C₆alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl.

Alternatively, A is OH; X is COCH₃; W, Y, and Z are as described above;and R⁴ is selected from the groups consisting of:

where R²¹ is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl, orC₁-C₄ alkheteroaryl, R²⁰ is H, C₁-C₆ alkyl, COR¹⁹, CO₂R¹⁹, or CONHR¹⁹,CSR₁₉, COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, SO₂NHR¹⁹, where R¹⁹ is C₁-C₆alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl.

Alternatively, A is H or OH; X is H or COCH₃; W, Y, and Z are asdescribed above; and R⁴ is

with the proviso that one or both of Y and Z are halogen.

Alternatively, A is H or OH; X is H or COCH₃; W, Y, and Z are asdescribed above; and R⁴ is

where R²² is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl,C₁-C₄ alkheteroaryl, COR²⁴, CO₂R²⁴, CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴,CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl,C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl, and r is 1-2.

Alternatively, A is H or OH; X is H or COCH₃; W, Y, and Z are asdescribed above; and R⁴ is

where R²¹ is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl, orC₁-C₄ alkheteroaryl.

Alternatively, A is H or OH; X is H or COCH₃; W, Y, and Z are asdescribed above; and R⁴ is

where ═E is ═O or (H,H), R²² is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl,C₁-C₄ alkaryl, C₁-C₄ alkheteroaryl, COR²⁴, CO₂R²⁴, CONHR²⁴, CSR₂₄,COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO²NHR²⁴, where R²⁴ is C₁-C₆ alkyl,C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl, r is1-2, and s is 0-1.

Alternatively, A is H or OH; X is H or COCH₃; W, Y, and Z are asdescribed above; and R⁴

where R²² is H C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, CONHR²⁴, CSR²⁴, COSR²⁴,CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, where R²⁴ is C₁-C₆ alkyl, C₆-C₁₂aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl.

In one embodiment, A is H or OH; X is H or COCH₃; W, Y, and Z are asdescribed above; and R⁴ is

where one or both of Y and Z is F.

In another embodiment, W is O; Y is H; Z is H; A is OH, X is H or COCH₃,and R⁴ is

where each of R⁵ and R⁶ is H, or R⁵ and R⁶ together are ═O, each of R⁸,R⁹, R¹², R¹³ and R¹⁵ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, each of R¹⁰and R¹¹ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R¹⁰ and R¹¹ together are═O, R¹⁷ is H, C₁-C₆ alkyl, COR¹⁹, CO₂R¹⁹, or CONHR¹⁹, CSR₁₉, COSR¹⁹,CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or SO₂NHR¹⁹, where R¹⁹ is C₁-C₆ alkyl, C₆-C₁₂aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl.

In another embodiment, W is O; Y is H; Z is H; A is H or OH, X is H orCOCH₃, and R⁴ is

In another embodiment, W is O; Y is H; Z is H; A is H or OH, X is H orCOCH₃, and R⁴ is

In another embodiment, W is O; Y is H; Z is H; X is H or COCH₃; A is Hor OH; and R⁴ is selected from the group consisting of:

where R²⁰ and R²¹ are as described above, or

W is O; Y is H; Z is H; X is H or COCH₃, A is H or OH; and R⁴is:

where each of R¹⁷ and R²³ is, independently, H, C₁-C₆ alkyl, COR²⁴,CO₂R²⁴, or CONHR²⁴, where R²⁴ is C₁-C₆ alkyl, C₁-C₄ alkaryl, heteroaryl,or C₁-C₄ alkheteroaryl., or

W is O, Y is H, Z is H, X is COCH₃, A is OH, and R⁴ is selected from thegroup consisting of

where R¹⁶ and R¹⁷ are as described above.

The invention also features pharmaceutical compositions that include acompound of formula (I) and a pharmaceutically acceptable carrier ordiluent.

In another aspect, the invention features a method of killing, treating,or preventing a microbial infection in an animal, preferably a mammal,and most preferably a human, that includes administering to the animal apharmaceutical composition of the invention. The invention furtherfeatures treating or preventing diseases associated with such microbialinfections. Such methods of treatment or prevention may include theoral, topical, intravenous, intramuscular, or subcutaneneousadministration of compositions of the invention.

The invention also features a method for treating or preventing thedevelopment of an atherosclerosis-associated disease in a patient byadministering to the patient a compound of formula (I) in an amounteffective to treat or prevent the development of theatherosclerosis-associated disease in the patient. The patient istypically diagnosed as having the atherosclerosis-associated disease (orbeing at increased risk of developing the disease) or as havingmacrophages or foam cells infected with C. pneumoniae prior to theadministration of a compound of formula (I).

The invention also features a method of reducing the level of C-reactiveprotein in a patient in need thereof by administering to the patient acompound of formula (I) in an amount effective to reduce the level ofC-reactive protein in the patient. In one embodiment, the patient hasnot been diagnosed as having a bacterial infection. In anotherembodiment, the patient has been diagnosed as having macrophages or foamcells infected with C. pneumoniae.

The invention also features a method for reducing C. pneumoniaereplication in macrophages or foam cells in a patient in need thereof byadministering a compound of formula (I) to the patient in an amounteffective to reduce C. pneumoniae replication in macrophages or foamcells in the patient.

The invention also features a method for treating a persistent C.pneumoniae infection in macrophages or foam cells in a patient byadministering a compound of formula (I) to the patient in an amounteffective to treat the C. pneumoniae infection in macrophages or foamcells in the patient.

The invention also features a method for treating a chronic diseaseassociated with an infection of C. pneumoniae by administering acompound of formula (I) to the patient in an amount effective to treatthe infection.

In any of the foregoing aspects, the dosage of a compound of formula (I)is normally about 0.001 to 1000 mg/day. The compound may be given daily(e.g., a single oral dose of 2.5 to 25 mg/day) or less frequently (e.g.,a single oral dose of 5, 12.5, or 25 mg/week). Treatment may be for oneday to one year, or longer. In one embodiment, a compound of formula (I)is administered at an initial does of 2.5 to 100 mg for one to sevenconsecutive days, followed by a maintenance dose of 0.005 to 10 mg onceevery one to seven days for one month, one year, or even for the life ofthe patient.

If desired, a compound of formula (I) may be administered in conjunctionwith one or more additional agents such as anti-inflammatory agents(e.g., non-steroidal anti-inflammatory drugs (NSAIDs; e.g., detoprofen,diclofenac, diflunisal, etodolac, fenoprofen, flurbiprofen, ibuprofen,indomethacin, ketoprofen, meclofenameate, mefenamic acid, meloxicam,nabumeone, naproxen sodium, oxaprozin, piroxicam, sulindac, tolmetin,celecoxib, rofecoxib, aspirin, choline salicylate, salsalte, and sodiumand magnesium salicylate) and steroids (e.g., cortisone, dexamethasone,hydrocortisone, methylprednisolone, prednisolone, prednisone,triamcinolone)), antibacterial agents (e.g., azithromycin,clarithromycin, erythromycin, gatifloxacin, levofloxacin, amoxicillin,metronidazole, penicillin G, penicillin V, methicillin, oxacillin,cloxacillin, dicloxacillin, nafcillin, ampicillin, carbenicillin,ticarcillin, mezlocillin, piperacillin, azlocillin, temocillin,cepalothin, cephapirin, cephradine, cephaloridine, cefazolin,cefamandole, cefuroxime, cephalexin, cefprozil, cefaclor, loracarbef,cefoxitin, cefmatozole, cefotaxime, ceftizoxime, ceftriaxone,cefoperazone, ceftazidime, cefixime, cefpodoxime, ceftibuten, cefdinir,cefpirome, cefepime, BAL5788, BAL9141, imipenem, ertapenem, meropenem,astreonam, clavulanate, sulbactam, tazobactam, streptomycin, neomycin,kanamycin, paromycin, gentamicin, tobramycin, amikacin, netilmicin,spectinomycin, sisomicin, dibekalin, isepamicin, tetracycline,chlortetracycline, demeclocycline, minocycline, oxytetracycline,methacycline, doxycycline, telithromycin, ABT-773, lincomycin,clindamycin, vancomycin, oritavancin, dalbavancin, teicoplanin,quinupristin and dalfopristin, sulphanilamide, para-aminobenzoic acid,sulfadiazine, sulfisoxazole, sulfamethoxazole, sulfathalidine,linezolid, nalidixic acid, oxolinic acid, norfloxacin, perfloxacin,enoxacin, ofloxacin, ciprofloxacin, temafloxacin, lomefloxacin,fleroxacin, grepafloxacin, sparfloxacin, trovafloxacin, clinafloxacin,moxifloxacin, gemifloxacin, sitafloxacin, daptomycin, garenoxacin,ramoplanin, faropenem, polymyxin, tigecycline, AZD2563, ortrimethoprim), platelet aggregation inhibitors (e.g., abciximab,aspirin, cilostazol, clopidogrel, dipyridamole, eptifibatide,ticlopidine, or tirofiban), anticoagulants (e.g., dalteparin,danaparoid, enoxaparin, heparin, tinzaparin, or warfarin), antipyretics(e.g., acetaminophen), or lipid lowering agents (e.g., cholestyramine,colestipol, nicotinic acid, gemfibrozil, probucol, ezetimibe, or statinssuch as atorvastatin, rosuvastatin, lovastatin simvastatin, pravastatin,cerivastatin, and fluvastatin). These additional agents may beadministered within 14 days, 7 days, 1 day, 12 hours, or 1 hour ofadministration of the rifamycin analog, or simultaneously therewith. Theadditional therapeutic agents may be present in the same or differentpharmaceutical compositions as the rifamycin analog. When present indifferent pharmaceutical compositions, different routes ofadministration may be used. For example, a compound of formula (I) maybe administered orally, while a second agent may be administered byintravenous, intramuscular, or subcutaneous injection.

The invention also features a stent coated with a compound of formula(I). The stent can be, e.g., a wire mesh tube used to hold open anartery. Stents are typically inserted following angioplasty.

The invention also features methods and compositions for treating orpreventing an ear infection in a patient by orally administering ortopically administering to the affected otic area (e.g., the tympanicmembrane or the external auditory canal of the ear) of the patient apharmaceutical composition including a therapeutically effective amountof a compound of formula (I). The compositions and methods of theinvention can also be used to treat or prevent infections that resultfrom surgery.

The invention also features a pharmaceutical composition suitable fortopical administration to the ear of a patient containing a compound offormula (I) and a pharmaceutically-acceptable excipient, administered ata dose capable of reducing the infection in the patient. According tothis invention, the compound of formula (I) can be in the amount between0.001% and 5% weight/volume (w/v), preferably 0.01% and 3% w/v, morepreferably 0.1% and 1% w/v, or most preferably 0.1% and 0.4% w/v. Thecompound of formula (I) can also be impregnated in a porous media (forexample, an ear wick such as a sponge, gauze, cotton, orhydrocellulose), which is suitable for insertion into the ear of apatient. If desired, the composition may also include one or morepenetration enhancers (e.g., alcohols, polyols, sulfoxides, esters,ketones, amides, oleates, surfactants, alkanoic acids, lactam compounds,alkanols, or admixtures thereof).

In another aspect, the invention also features a method for treating orpreventing the development of an ear infection in a patient using acomposition described above. A compound of formula (I) can beadministered to the infected ear by means of drops or by the insertionof a rifamycin analog-impregnated porous media into the external earcanal to the tympanic membrane. Ear infections that can be treated usingthe methods and composition of the invention include otitis media andotitis externa. Types of otitis media amenable to treatment include, forexample, acute otitis media, otitis media with effusion, and chronicotitis media. Types of otitis externa include acute otitis externa,chronic otitis externa, and malignant otitis externa. A rifamycin analogof the invention is administered to the ear (e.g., the tympanic membraneor the external auditory canal of the ear) to treat or prevent bacterialinfections associated with otitis media (e.g., an infection of H.influenza, M. catarhalis, or S. pneumoniae) or in otitis externa (e.g.,an infection of S. intermedius, Streptococcus spp. Pseudomonas spp.,Proteus spp., or E. coli).

The methods and compositions of the invention are also useful to treatinfections associated with otic surgical procedures such astympanoplasty, stapedectomy, removal of tumors, or cochlear implantsurgery. The compositions may also be used prophylactically, prior totherapies or conditions that can cause ear infections. Compositionscontaining a compound of formula (I) can therefore be applied to an areaof the ear to which the surgical intervention will be performed, withinat least seven days (before or after) of the surgical intervention. Whentreating a patient affected with otitis externa, an acidificationtherapy involving the administration of an acetic acid solution to theear of the patient may also be performed.

Typically, patients are administered one to four drops of a compound ofthe invention in a total amount between 0.001% and 5% w/v, preferably0.01% and 3% w/v, more preferably 0.1% and 1% w/v, or most preferably0.1% and 0.4% w/v. The composition may be given daily (e.g., once,twice, three times, or four times daily) or less frequently (e.g., onceevery other day, or once or twice weekly). Treatment may be for 1 to 21days, desirably 1 to 14 days, or even 3 to 7 days. Additionaltherapeutic agents, such as anti-inflammatory agents (e.g.,non-steroidal anti-inflammatory or steroid), anesthetics, zinc salts, orother antimicrobial agents, can also be administered with a rifamycinanalog of the invention. Non-steroidal anti-inflammatory agents include,for example, detoprofen, diclofenac, diflunisal, etodolac, fenoprofen,flurbiprofen, indomethacin, ketoprofen, mechlofenameate, mefenamic acid,meloxicam, nabumeone, naproxen sodium, oxaprozin, piroxicam, sulindac,tolmeting, celecoxib, rofecoxib, choline salicylate, salsate, sodiumsalicylate, magnesium salicylate, aspirin, ibuprofen, paracetamol,acetaminophen, and pseudoephedrine and steroids include, for example,hydrocortisone, prednisone, fluprednisolone, triamcinolone,dexamethasone, betamethasone, cortisone, prednilosone,methylprednisolone, fluocinolone acetonide, flurandrenolone acetonide,and fluorometholone. Anesthetics according to the invention can be, forexample, benzocaine, butamben picrate, tetracaine, dibucaine,prilocaine, etidocaine, mepivacaine, bupivicaine, and lidocaine. A zincsalt can be zinc sulfate, zinc chloride, zinc acetate, zinc phenolsulfonate, zinc borate, zinc bromide, zinc nitrate, zincglycerophosphate, zinc benzoate, zinc carbonate, zinc citrate, zinchexafluorosilicate, zinc diacetate trihydrate, zinc oxide, zincperoxide, zinc salicylate, zinc silicate, zinc stannate, zinc tannate,zinc titanate, zinc tetrafluoroborate, zinc gluconate, and zincglycinate, and antimicrobial agents according to the invention include,for example, azithromycin, clarithromycin, erythromycin, gatifloxacin,levofloxacin, amoxicillin, metronidazole, penicillin G, penicillin V,methicillin, oxacillin, cloxacillin, dicloxacillin, nafcillin,ampicillin, carbenicillin, ticarcillin, mezlocillin, piperacillin,azlocillin, temocillin, cepalothin, cephapirin, cephradine,cephaloridine, cefazolin, cefamandole, cefuroxime, cephalexin,cefprozil, cefaclor, loracarbef, cefoxitin, cefmatozole, cefotaxime,ceftizoxime, ceftriaxone, cefoperazone, ceftazidime, cefixime,cefpodoxime, ceftibuten, cefdinir, cefpirome, cefepime, BAL5788,BAL9141, imipenem, ertapenem, meropenem, astreonam, clavulanate,sulbactam, tazobactam, streptomycin, neomycin, kanamycin, paromycin,gentamicin, tobramycin, amikacin, netilmicin, spectinomycin, sisomicin,dibekalin, isepamicin, tetracycline, chlortetracycline, demeclocycline,minocycline, oxytetracycline, methacycline, doxycycline, telithromycin,ABT-773, lincomycin, clindamycin, vancomycin, oritavancin, dalbavancin,teicoplanin, quinupristin and dalfopristin, sulphanilamide,para-aminobenzoic acid, sulfadiazine, sulfisoxazole, sulfamethoxazole,sulfathalidine, linezolid, nalidixic acid, oxolinic acid, norfloxacin,perfloxacin, enoxacin, ofloxacin, ciprofloxacin, temafloxacin,lomefloxacin, fleroxacin, grepafloxacin, sparfloxacin, trovafloxacin,clinafloxacin, moxifloxacin, gemifloxacin, sitafloxacin, daptomycin,garenoxacin, ramoplanin, faropenem, polymyxin, tigecycline, AZD2563, ortrimethoprim. These additional therapeutic agents can be present in thesame or different pharmaceutical compositions as the rifamycin analog offormula (I). When a therapeutic agent is present in a differentpharmaceutical composition, different routes of administration may beused. The rifamycin analog of formula (I) and the second therapeuticagent, for example, may also be administered within 24 hours of eachother, and an anti-inflammatory agent, for example, may be administeredorally, or by intravenous, intramuscular, or subcutaneous injection.

To increase the efficacy of the topically administered rifamycinanalog-containing composition, it is desirable that the amount of debrisand granulation tissue are reduced in the infected ear of the patient atleast one hour prior to the administration of the rifamycin and at leastonce a day. Debris can be removed, for example, by suction, irrigationwith a solution containing hydrogen peroxide, cauterization, or bymanual techniques employing microinstruments and microscope. Reductionin the amount of granulation tissue in the infected ear may be performedby means of cautering, or by the administration of a steroid.

The invention also features a pharmaceutical pack containing (i) acompound of formula (I) in an amount effective to treat a patient havingan ear infection; and (ii) instructions for administering the compoundto the ear of a patient. The invention also features a containercontaining a rifamycin analog of formula (I) and a pharmaceuticalexcipient suitable for topical administration to the ear. If desired, anapplicator for applying the composition to the ear may also be included.Desirably, the rifamycin analog can be in the amount between 0.001% and5% weight/volume (w/v), preferably 0.01% and 3% w/v, more preferably 0.1% and 1% w/v, or most preferably 0.1% and 0.4% w/v and is present inamounts sufficient to treat for at least 1, 3, 5, 7, 10, 14, or 21 days.A penetration enhancer may also be added (e.g., alcohols, polyols,sulfoxides, esters, ketones, amides, oleates, surfactants, alkanoicacids, lactam compounds, alkanols, or admixtures thereof).

The invention also features a method for treating chronic gastritis,gastric ulcer, or duodenal ulcer associated with an infection of H.pylori, or preventing the disease or infection, in a patient. The methodincludes the step of orally administering to the patient an effectiveamount of a compound of formula (I) to treat the patient. The compoundis normally about 0.1 to 1000 mg/day (desirably about 1 to 100 mg/day,more desirably about 1 to 50 mg/day, and even more desirably about 1 to25 mg/day). The compound may be given daily (e.g., once, twice, threetimes, or four times daily) or less frequently (e.g., once every otherday, or once or twice weekly). Treatment may be for 1 to 21 days,desirably 1 to 14 days or even 3 to 7 days. If desirable, a rifamycinanalog can be administered with a proton pump inhibitor (e.g.,omeprazole, esomeprazole, lansoprazole, leminoprazole, pantoprazole orrobeprazole), and/or bismuth preparation (e.g., colloidal bismuthsubcitrate or bismuth subsalicylate).

The invention also features a pharmaceutical pack including (i) acompound of formula (I) in an amount effective to treat chronicgastritis, gastric ulcer, or duodenal ulcer associated with an infectionof H. pylori in a patient; and (ii) instructions for administering thecompound to the patient. Desirably, the compound is in unit amounts ofbetween 0.1 and 1000 mg (e.g., between 1 and 50 mg or between 5 and 50mg), and is present in amounts sufficient to treat for at least 1, 3, 5,7, 10, 14, or 21 days. The pack may optionally include a proton pumpinhibitor and/or bismuth preparation. In one embodiment, the rifamycinanalog of formula (I) is in a pharmaceutical composition with the protonpump inhibitor and/or bismuth preparation.

The invention also features a method for treating a patient havingantibiotic-associated bacterial diarrhea or an infection of C.difficile, or preventing the disease or infection in the patient. Themethod includes the step of orally administering to the patient aneffective amount of a compound of formula (I) to treat the patient. Thecompound is normally about 0.1 to 1000 mg/day (desirably about 1 to 100mg/day, more desirably about 1 to 50 mg/day, and even more desirablyabout 1 to 25 mg/day). The compound may be given daily (e.g., once,twice, three times, or four times daily) or less frequently (e.g., onceevery other day, or once or twice weekly). Treatment may be for 1 to 21days, desirably 1 to 14 days or even 3 to 7 days. In one embodiment, arifamycin analog is administered at an initial dose of between 5 and 100mg, followed by subsequent doses of between 1 and 50 mg for 3 to 7 days.A single dose (e.g., in a dosage of between 5 and 50 mg) can also beemployed in the method of the invention. If desirable, a rifamycinanalog of formula (I) can be administered with a second antibiotic(e.g., metronidazole or vancomycin), either simultaneously orsequentially.

The invention also features a pharmaceutical pack including (i) acompound of formula (I) in an amount effective to treat a patient havingantibiotic-associated bacterial diarrhea or an infection of C.difficile; and (ii) instructions for administering the compound to thepatient for treating or preventing a C. difficile infection. Desirably,the compound is in unit amounts of between 1 and 1000 mg (e.g., between1 and 50 mg or between 5 and 50 mg), and is present in amountssufficient to treat for at least 1, 3, 5, 7, 10, 14, or 21 days.

The invention features a method for treating a patient having aninfection of Chlamydia trachomatis. The method includes the step ofadministering to the patient a compound of formula (I) in an amounteffective to treat the patient. In one embodiment, the patient isadministered a single oral dose of a compound of formula (I).

The invention also features a pharmaceutical pack that includes (i) asingle oral dose of a compound of formula (I) in an amount effective totreat a patient having an infection of C. trachomatis or N. gonorrhoeae;and (ii) instructions for administering the single oral dose to thepatient. Desirably, the dose is in an amount between 0.1 and 100 mg(e.g., between 1 and 50 mg or between 5 and 25 mg).

The invention also features a method of treating a patient having achronic disease associated with a bacterial infection caused by bacteriacapable of establishing a cryptic phase. This method includes the stepof administering to a patient a compound of formula (I) for a time andin an amount sufficient to treat the cryptic phase of the bacterialinfection. The chronic disease may be an inflammatory disease. Examplesof inflammatory diseases include but are not limited to asthma, coronaryartery disease, arthritis, conjunctivitis, lymphogranuloma venerum(LGV), cervicitis, and salpingitis. The chronic disease can also be anautoimmune disease (e.g., systemic lupus erythematosus, diabetesmellitus, or graft versus host disease).

The invention also features a method for treating a patient diagnosed asbeing infected with a bacterium having a multiplying form and anon-multiplying form by administering to the patient (i) a rifamycinanalog of formula (I) and (ii) a second antibiotic that is effectiveagainst the multiplying form of the bacterium, wherein the twoantibiotics are administered in amounts and for a duration that, incombination, treat the patient.

In a related aspect, the invention features a method of treating achronic disease associated with a persistent bacterial infection ortreating the persistent bacterial infection itself by administering acompound of formula (I).

In preferred embodiments of any of the foregoing aspects, the persistentintracellular bacterial infection is caused by one of the following:Chlamydia spp. (e.g., C. trachomatis, C. pneumoniae, C. psittaci, C.suis, C. pecorum, C. abortus, C. caviae, C. felis, C. muridarum), N.hartmannellae, W. chondrophila, S. negevensis, or P. acanthamoeba.

The time sufficient to treat a bacterial infection ranges from one weekto one year, but it can also be extended over the lifetime of theindividual patient, if necessary. In more preferable embodiments, theduration of treatment is at least 30 days, at least 45 days, at least 10days, or at least 180 days. Ultimately, it is most desirable to extendthe treatment for such a time that the bacterial infection is no longerdetected.

The rifamycin analogs of formula (I) are useful against drug resistantGram-positive cocci such as methicillin-resistant S. aureus andvancomycin-resistant enterococci, and are useful in the treatment ofcommunity-acquired pneumonia, upper and lower respiratory tractinfections, skin and soft tissue infections, hospital-acquired lunginfections, bone and joint infections, and other bacterial infections.

The compounds and methods of the present invention can be used to treat,for example, respiratory tract infections, acute bacterial otitis media,bacterial pneumonia, urinary tract infections, complicated infections,noncomplicated infections, pyelonephritis, intra-abdominal infections,deep-seated abcesses, bacterial sepsis, skin and skin structureinfections, soft tissue infections, bone and joint infections, centralnervous system infections, bacteremia, wound infections, peritonitis,meningitis, infections after burn, urogenital tract infections,gastrointestinal tract infections, pelvic inflammatory disease,endocarditis, and other intravascular infections.

The compounds and methods of the present invention can also be used totreat diseases associated with bacterial infection. For example,bacterial infections can produce inflammation, resulting in thepathogenesis of atherosclerosis, multiple sclerosis, rheumatoidarthritis, diabetes, Alzheimer's disease, asthma, cirrhosis of theliver, psoriasis, meningitis, cystic fibrosis, cancer, or osteoporosis.Accordingly, the present invention also features a method of treatingthe diseases associated with bacterial infection listed above.

The methods of the present invention can be used to treat or preventinfections by bacteria from a variety of genera, such as Escherichiaspp., Enterobacter spp., Enterobacteriaceae spp., Klebsiella spp.,Serratia spp., Pseudomonas spp., Acinetobacter spp., Bacillus spp.,Micrococcus spp., Arthrobacter spp., Peptostreptococcus spp.,Staphylococcus spp., Enterococcus spp., Streptococcus spp., Haemophilusspp., Neisseria spp., Bacteroides spp., Citrobacter spp., Branhamellaspp., Salmonella spp., Shigella spp., Proteus spp., Clostridium spp.,Erysipelothrix spp., Listeria spp., Pasteurella spp., Streptobacillusspp., Spirillum spp., Fusospirocheta spp., Treponema spp., Borreliaspp., Actinomycetes spp., Mycoplasma spp., Chlamydia spp., Rickettsiaspp., Spirochaeta spp., Legionella spp., Mycobacteria spp., Ureaplasmaspp., Streptomyces spp., and Trichomoras spp. Accordingly, the inventionfeatures a method of treating infections by the bacteria belonging tothe genera above, among others.

Particular Gram-positive bacterial infections that can be treatedaccording to the method of the invention include infections byStaphylococcus aureus, Staphylococcus epidermidis, Enterococcusfaecalis,Enterococcus faecium, Clostridium perfringens, Streptococcus pyogenes,Streptococcus pneumoniae, other Streptococcus spp., and otherClostridium spp.

Multi-drug resistant strains of bacteria can be treated according to themethods of the invention. Resistant strains of bacteria includepenicillin-resistant, methicillin-resistant, quinolone-resistant,macrolide-resistant, and/or vancomycin-resistant bacterial strains. Themulti-drug resistant bacterial infections to be treated using themethods of the present invention include infections by penicillin-,methicillin-, macrolide-, vancomycin-, and/or quinolone-resistantStreptococcus pneumoniae; penicillin-, methicillin-, macrolide-,vancomycin-, and/or quinolone-resistant Staphylococcus aureus;penicillin-, methicillin-, macrolide-, vancomycin-, and/orquinolone-resistant Streptococcus pyogenes; and penicillin-,methicillin-, macrolide-, vancomycin-, and/or quinolone-resistantenterococci.

The invention also features a method of eradicating non-multiplyingbacteria not eradicated in a patient following treatment with a firstantibiotic by administering to the patient a rifamycin antibiotic offormula (I) or (II) in an amount and for a duration sufficient toeradicate the non-multiplying bacteria in the patient.

Compounds of the invention may also be used to treat or prevent viralinfections.

In another aspect, the invention features a pharmaceutical compositionthat includes a compound described herein in any pharmaceuticallyacceptable form, including isomers such as diastereomers andenantiomers, salts, solvates, and polymorphs thereof. In variousembodiments, the composition includes a compound of the invention alongwith a pharmaceutically acceptable carrier or diluent.

In another aspect, the invention features a method of treating amicrobial infection in an animal comprising co-administering a compoundof the invention along with one or more antifungal agents, antiviralagents, antibacterial agents, or antiprotozoan agents, or combinationsthereof.

In any of the above aspects, desirable rifamycin analogs of formula (I)include 4′-fluoro-5′-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin,4′-fluoro-5′-(1-piperazinyl)benzoxazinorifamycin,4′-fluoro-5′-(3-methyl-1-piperazinyl)benzoxazinorifamycin,4′-methoxy-6′-fluoro-5′-(3-methyl-1-piperazinyl)benzoxazinorifamycin,4′,6′-difluoro-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzoxazinorifamycin,4′-fluoro-6′-methoxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,4′-fluoro-5′-[6-amino-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(3-methyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-4′-methoxy-6′-fluoro-5′-(3-methyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-4′,6′-difluoro-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzoxazinorifamycin,25-O-deacetyl-4′-fluoro-6′-methoxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-4′-fluoro-5′-[6-amino-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-5′-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-fluoro-5′-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-fluoro-5′-(1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-fluoro-5′-(3-methyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-methoxy-6′-fluoro-5′(3-methyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′,6′-difluoro-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-fluoro-6′-methoxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-fluoro-5′-[6-amino-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,4′-fluoro-5′-(4-isobutyl-1-piperazinyl)benzthiazinorifamycin,4′-fluoro-5′-(1-piperazinyl)benzthiazinorifamycin,4′-fluoro-5′-(3-methyl-1-piperazinyl)benzthiazinorifamycin,4′-methoxy-6′-fluoro-5′-(3-methyl-1-piperazinyl)benzthiazinorifamycin,4′,6′-difluoro-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin,4′-fluoro-6′-methoxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzthiazinorifamycin,4′-fluoro-5′-[6-amino-3-azabicyclo[3.1.0]hex-3-yl]benzthiazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(4-isobutyl-1-piperazinyl)benzthiazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(1-piperazinyl)benzthiazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(3-methyl-1-piperazinyl)benzthiazinorifamycin,25-O-deacetyl-4′-methoxy-6′-fluoro-5′-(3-methyl-1-piperazinyl)benzthiazinorifamycin,25-O-deacetyl-4′,6′-difluoro-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin,25-O-deacetyl-4′-fluoro-6′-methoxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzthiazinorifamycin,25-O-deacetyl-4′-fluoro-5′-[6-amino-3-azabicyclo[3.1.0]hex-3-yl]benzthiazinorifamycin,3′-hydroxy-5′-((3R,5S)-3,5-dimethylpiperazinyl)benzoxazinorifamycin,3′-hydroxy-5′-((3R,5S)-3,5-diethylpiperazinyl)benzoxazinorifamycin,3′-hydroxy-5′-((3R,5S)-3-ethyl-5-methylpiperazinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-((3R,5S)-3,5-dimethylpiperazinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-((3R,5S)-3-ethyl-5-methylpiperazinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-((3R,5S)-3,5-diethylpiperazinyl)benzoxazinorifamycin,3′-hydroxy-5′-((4aR,7aR)octahydro-1H-pyrrolyl[3,4-b]pyridine)benzoxazinorifamycin,3′-hydroxy-5′-((4aS,7aS)octahydro-1H-pyrrolyl[3,4-b]pyridine)benzoxazinorifamycin,3′-hydroxy-5′-((8aR)-octahydropyrrolyl[1,2-a]pyrazine)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-((8aR)-octahydropyrrolyl[1,2-a]pyrazine)benzoxazinorifamycin,3′-hydroxy-5′-((8aS)-octahydropyrrolyl[1,2-a]pyrazine)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-((8aS)-octahydropyrrolyl[1,2-a]pyrazine)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-(4-methylpiperazinyl)benzoxazinorifamycin,3′-hydroxy-5′-(ethyl piperidinyl-4-ylcarbamate)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-(ethylpiperidinyl-4-ylcarbamate)benzoxazinorifamycin,3′-hydroxy-5′-((3Z)-4-(aminomethyl)pyrrolidinyl-3-one O-methyloxime)benzoxazinorifamycin, 3′-hydroxy-5′-(5-azaspiro[2.4]heptan-7-amino-5-yl)benzoxazinorifamycin, 3′-hydroxy-5′-(5-aminopyrrolidinyl)benzoxazinorifamycin,3′-hydroxy-5′-(4-ethylcarbamyl-1-piperidinyl)benzoxazinorifamycin(compound 1),3′-hydroxy-5′-[6-(2-trimethylsilyl)ethylcarbamyl1-(1R,5S)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin(compound 2),25-O-deacetyl-3′-hydroxy-5′-(4-ethylcarbamyl-1-piperidinyl)benzoxazinorifamycin(compound 3),3′-hydroxy-5′-[6-amino-(1R,5S)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin(compound 4),3′-hydroxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin(compound 5), 3′-hydroxy-5′-(1-piperidinyl-4-(N-phenyl)propanamide)benzoxazinorifamycin (compound 6),25-O-deacetyl-3′-hydroxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin(compound 7),25-O-deacetyl-3′-hydroxy-5′-(1-piperidinyl-4-(N-phenyl)propanamide)benzoxazinorifamycin(compound 8),3′-hydroxy-5′-(4-morpholinyl-1-piperidinyl)benzoxazinorifamycin(compound 9),3′-hydroxy-5′-(3,8-diazabicyclo[3.2.1]octan-3-yl)benzoxazinorifamycin(compound 10),25-O-deacetyl-3′-hydroxy-5′-(4-morpholinyl-1-piperidinyl)benzoxazinorifamycin(compound 11),3′-hydroxy-5′-[(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin(compound 12),3′-hydroxy-5′-(4-(2-methylpropyl)carbamyl-1-piperidinyl)benzoxazinorifamycin(compound 13),25-O-deacetyl-3′-hydroxy-5′-(4-(2-methylpropyl)carbamyl-1-piperidinyl)benzoxazinorifamycin(compound 14),25-O-deacetyl-3′-hydroxy-5′-[(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin(compound 15),25-O-deacetyl-3′-hydroxy-5′-(3,8-diazabicyclo[3.2.1]octan-3-yl)benzoxazinorifamycin(compound 16),3′-hydroxy-5′-(4-N,N-dimethylamino-1-piperidinyl)benzoxazinorifamycin(compound 17), 25-O-deacetyl-3′-hydroxy-5′-(4-N,Ndimethylamino-1-piperidinyl)benzoxazinorifamycin (compound 18),5′-(4-ethylcarbamyl-1-piperidinyl)-N′-methylbenzodiazinorifamycin(compound 19),25-O-deacetyl-3′-hydroxy-5′-[6-amino-(1R,5S)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin(compound 20),3′-hydroxy-5′-[6-ethylcarbamyl-(1R,5s)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin(compound 21),3′-hydroxy-5′-[4-isopropylcarbamyl-1-piperidinyl]benzoxazinorifamycin(compound 22),3′-hydroxy-5′-[4-trifluoromethylsulfonyl-1-piperidinyl]benzoxazinorifamycin(compound 23),3′-hydroxy-5′-[4-butanamide-1-piperidinyl]benzoxazinorifamycin (compound24), 3′-hydroxy-5′-[4-methylsulfonyl-1-piperidinyl]benzoxazinorifamycin(compound 25),25-O-deacetyl-3′-hydroxy-5′-[4-propyluryl-1-piperidinyl]benzoxazinorifamycin(compound 26),25-O-deacetyl-3′-hydroxy-5′-[4-methylsulfonyl-1-piperidinyl]benzoxazinorifamycin(compound 27),3′-hydroxy-5′-[4-propyluryl-1-piperidinyl]benzoxazinorifamycin (compound28),25-O-deacetyl-3′-hydroxy-5′-[4-isopropylcarbamyl-1-piperidinyl]benzoxazinorifamycin(compound 29),25-O-deacetyl-3′-hydroxy-5′-[4-methylcarbamyl-1-piperidinyl]benzoxazinorifamycin(compound 30),25-O-deacetyl-5′-(4-ethylcarbamyl-1-piperidinyl)-N′-methylbenzdiazinorifamycin(compound31),3′-hydroxy-5′-[4-methylcarbamyl-1-piperidinyl]benzoxazinorifamycin(compound 32),3′-hydroxy-5′-[4-amino-1-piperidinyl]benzoxazinorifamycin(compound33),3′-hydroxy-5′-[4-ethyluryl-1-piperidinyl]benzoxazinorifamycin(compound34),3′-hydroxy-5′-[4-propylsulfonyl-1-piperidinyl]benzoxazinorifamycin(compound 35),25-O-deacetyl-3′-hydroxy-5′-[4-butanamide-1-piperidinyl]benzoxazinorifamycin(compound 36),25-O-deacetyl-3′-hydroxy-5′-[4-ethyluryl-1-piperidinyl]benzoxazinorifamycin(compound 37),25-O-deacetyl-3′-hydroxy-5′-[4-trifluoromethysulfonyl-1-piperidinyl]benzoxazinorifamycin(compound 38),25-O-deacetyl-3′-hydroxy-5′-[4-amino-1-piperidinyl]benzoxazinorifamycin(compound39),3′-hydroxy-5′-[1-ethylcarbamyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin(compound40),3′-hydroxy-5′-[1-ethylcarbamyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,3′-hydroxy-5′-[4-methoxyethylcarbamyl-1-piperidinyl]benzoxazinorifamycin(compound 41),25-O-deacetyl-3′-hydroxy-5′-[1-ethylcarbamyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin(compound 42),25-O-deacetyl-3′-hydroxy-5′-[1-ethylcarbamyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-acetamide-1-piperidinyl]benzoxazinorifamycin(compound 43),3′-hydroxy-5′-[4-acetyl-1-piperidinyl]benzoxazinorifamycin(compound 44),3′-hydroxy-5′-[4-S-methylthiocarbamyl-1-piperidinyl]benzoxazinorifamycin(compound 45),25-O-deacetyl-3′-hydroxy-5′-[1-acetyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin(compound 46),25-O-deacetyl-3′-hydroxy-5′-[1-acetyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,3′-hydroxy-5′-[-acetyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin(compound 47),3′-hydroxy-5′-[1-acetyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,3′-hydroxy-5,′-[4-(2,2-dimethylethyl)carbamyl-1-piperidinyl]benzoxazinorifamycin(compound 48),3′-hydroxy-5′-[4-(4-(S-methylthiocarbamyl)-1-piperidinylcarbonyl)amino-1-piperidinyl]benzoxazinorifamycin(compound 49),3′-hydroxy-5′-[4-(4-methylpiperazinylcarbonyl)amino-1-piperidinyl]benzoxazinorifamycin(compound 50),3′-hydroxy-5′-[4-ethylcarbamylmethyl-1-piperidinyl]benzoxazinorifamycin(compound 51),25-O-deacetyl-3′-hydroxy-5′-[4-(2,2-dimethylethyl)carbamyl-1-piperidinyl]benzoxazinorifamycin(compound 52),3′-hydroxy-5′-[6-N,N-dimethylamino-(1R,5S)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin(compound 53),3′-hydroxy-5′-[6-N,N-dimethylamino-(1R,5S)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin(compound 54),3′-hydroxy-5′-[4-acetylaminomethyl-1-piperidinyl]benzoxazinorifamycin(compound 55),25-O-deacetyl-3′-hydroxy-5′-[4-acetylaminomethyl-1-piperidinyl]benzoxazinorifamycin(compound 56),3′-hydroxy-5′-[4-phenyl-1-piperidinyl]benzoxazinorifamycin (compound57),3′-hydroxy-5′-[1-methyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin(compound 58),3′-hydroxy-5′-[1-methyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[1-methyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin(compound 59),25-O-deacetyl-3′-hydroxy-5′-[1-methyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-ethylcarbamylmethyl-1-piperidinyl]benzoxazinorifamycin(compound 60),3′-hydroxy-5′-[4-(2-hydroxyethyl)-1-piperidinyl]benzoxazinorifamycin(compound 61),25-O-deacetyl-3′-hydroxy-5′-[4-phenyl-1-piperidinyl]benzoxazinorifamycin(compound 62),25-O-deacetyl-3′-hydroxy-5′-[4-methoxyethylcarbamyl-1-piperidinyl]benzoxazinorifamycin(compound 63),5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin (compound64), 5′-[(3S,SR)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin,25-O-deacetyl-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin(compound 65),25-O-deacetyl-5′-[(3S,5R)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-(2-hydroxyethyl)-1-piperidinyl]benzoxazinorifamycin(compound 66),25-O-deacetyl-3′-hydroxy-5′-[4-propylsulfonyl-1-piperidinyl]benzoxazinorifamycin(compound 67),5′-[(2S,5R)-4-(cyclopropylmethyl)-2,5-dimethylpiperazinyl]benzthiazinorifamycin(compound 68),5′-[(2R,5S)-4-(cyclopropylmethyl)-2,5-dimethylpiperazinyl]benzthiazinorifamycin,5′-[4-N,N-dimethylamino-1-piperidinyl]benzthiazinorifamycin (compound69),25-O-deacetyl-5′-[(2S,5R)-4-(cyclopropylmethyl)-2,5-dimethylpiperazinyl]benzthiazinorifamycin(compound 70),25-O-deacetyl-5′-[(2R,5S)-4-(cyclopropylmethyl)-2,5-dimethylpiperazinyl]benzthiazinorifamycin,3′-hydroxy-5′-[4-methyl-4-N,N-dimethylamino-1-piperidinyl]benzoxazinorifamycin(compound 71),3′-hydroxy-5′-[4-methyl-4-acetylamino-1-piperidinyl]benzoxazinorifamycin(compound 72),25-O-deacetyl-3′-hydroxy-5′-[4-methyl-4-N,N-dimethylamino-1-piperidinyl]benzoxazinorifamycin(compound 73),25-O-deacetyl-3′-hydroxy-5′-[4-methyl-4-acetylamino-1-piperidinyl]benzoxazinorifamycin(compound 74),3′-hydroxy-5′-[(3R)-N,N-dimethylamino-1-pyrrolidinyl]benzoxazinorifamycin(compound 75),3′-hydroxy-5′-[(3S)-N,N-dimethylamino-1-pyrrolidinyl]benzoxazinorifamycin,5′-[(8aS)octahydropyrrolo[1,2-a]pyrazin-2-yl]benzthiazinorifamycin(compound 76),5′-[(8aR)octahydropyrrolo[1,2-a]pyrazin-2-yl]benzthiazinorifamycin,25-O-deacetyl-5′-[(8aS)octahydropyrrolo[1,2-a]pyrazin-2-yl]benzthiazinorifamycin(compound 77),25-O-deacetyl-5′-[(8aR)octahydropyrrolo[1,2-a]pyrazin-2-yl]benzthiazinorifamycin(compound 78), or25-O-deacetyl-3′-hydroxy-5′-[3-hydroxy-1-azetidinyl]benzoxazinorifamycin(compound 79).

As used herein, the terms “alkyl” and the prefix “alk-” are inclusive ofboth straight chain and branched chain saturated or unsaturated groups,and of cyclic groups, i.e., cycloalkyl and cycloalkenyl groups. Unlessotherwise specified, acyclic alkyl groups are from 1 to 6 carbons.Cyclic groups can be monocyclic or polycyclic and preferably have from 3to 8 ring carbon atoms. Exemplary cyclic groups include cyclopropyl,cyclopentyl, cyclohexyl, and adamantyl groups. Alkyl groups may besubstituted with one or more substituents or unsubstituted. Exemplarysubstituents include alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio,halogen, alkylsilyl, hydroxyl, fluoroalkyl, perfluoralkyl, amino,aminoalkyl, disubstituted amino, quaternary amino, hydroxyalkyl,carboxyalkyl, and carboxyl groups. When the prefix “alk” is used, thenumber of carbons contained in the alkyl chain is given by the rangethat directly precedes this term, with the number of carbons containedin the remainder of the group that includes this prefix definedelsewhere herein. For example, the term “C₁-C₄ alkaryl” exemplifies anaryl group of from 6 to 18 carbons attached to an alkyl group of from 1to 4 carbons.

By “aryl” is meant a carbocyclic aromatic ring or ring system. Unlessotherwise specified, aryl groups are from 6 to 18 carbons. Examples ofaryl groups include phenyl, naphthyl, biphenyl, fluorenyl, and indenylgroups.

By “heteroaryl” is meant an aromatic ring or ring system that containsat least one ring hetero-atom (e.g., O, S, Se, N, or P). Unlessotherwise specified, heteroaryl groups are from 1 to 9 carbons.Heteroaryl groups include furanyl, thienyl, pyrrolyl, imidazolyl,pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl,tetrazolyl, oxadiazolyl, oxatriazolyl, pyridyl, pyridazyl, pyrimidyl,pyrazyl, triazyl, benzofuranyl, isobenzofuranyl, benzothienyl, indole,indazolyl, indolizinyl, benzisoxazolyl, quinolinyl, isoquinolinyl,cinnolinyl, quinazolinyl, naphtyridinyl, phthalazinyl, phenanthrolinyl,purinyl, and carbazolyl groups.

By “heterocycle” is meant a non-aromatic ring or ring system thatcontains at least one ring heteroatom (e.g., O, S, Se, N, or P). Unlessotherwise specified, heterocyclic groups are from 2 to 9 carbons.Heterocyclic groups include, for example, dihydropyrrolyl,tetrahydropyrrolyl, piperazinyl, pyranyl, dihydropyranyl,tetrahydropyranyl, dihydrofuranyl, tetrahydrofuranyl, dihydrothiophene,tetrahydrothiophene, and morpholinyl groups.

Aryl, heteroaryl, or heterocyclic groups may be unsubstituted orsubstituted by one or more substituents selected from the groupconsisting of C₁₋₆ alkyl, hydroxy, halo, nitro, C₁₋₆ alkoxy, C₁₋₆alkylthio, trifluoromethyl, C₁₋₆ acyl, arylcarbonyl, heteroarylcarbonyl,nitrile, C₁₋₆ alkoxycarbonyl, alkaryl (where the alkyl group has from 1to 4 carbon atoms) and alkheteroaryl (where the alkyl group has from 1to 4 carbon atoms).

By “alkoxy” is meant a chemical substituent of the formula —OR, where Ris an alkyl group. By “aryloxy” is meant a chemical substituent of theformula —OR′, where R′ is an aryl group. By “alkaryl” is meant achemical substituent of the formula —RR′, where R is an alkyl group andR′ is an aryl group, By “alkheteraryl” is meant a chemical substituentof the formula RR″, where R is an alkyl group and R″ is a heteroarylgroup.

By “halide” or “halogen” or “halo” is meant bromine, chlorine, iodine,or fluorine.

By “non-vicinal O, S, or NR” is meant an oxygen, sulfur, or nitrogenheteroatom substituent in a linkage, where the heteroatom substituentdoes not form a bond to a saturated carbon that is bonded to anotherheteroatom.

In structural representations where the chirality of a carbon has beenleft unspecified, it is to be presumed by one skilled in the art thateither chiral form of that stereocenter is possible.

By “benzoxazinorifamycin” is meant a compound described by formula (A):

where W is O. By “benzthiazinorifamycin” is meant a compound describedby formula (A), where W is S. By “benzdiazinorifamycin” is meant acompound described by formula (A), where W is N-R. Forbenzdiazinorifamycin, R can be H or an alkyl substituent. When R is analkyl substituent, it is referred to as N′-R (e.g., N′-methyl) in thenaming of the compound. Benzoxazinorifamycin, benzthiazinorifamycin, andbenzdiazinorifamycin analogs that contain substituents are numberedaccording to the numbering provided in formula (A). By “25-O-deacetyl”rifamycin is meant a rifamycin analog in which the acetyl group at the25-position has been removed. Analogs in which this position is furtherderivatized are referred to as a“25-O-deacetyl-25-(substituent)rifamycin”, in which the nomenclature forthe derivatizing group replaces “substituent” in the complete compoundname. For example, a benzoxazinorifamycin analog in which the25-acetyloxy group has been transformed to a carbonate group, with theother side of the carbonate bonded to a 2,3-dihydroxypropyl group, isreferred to as a “25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-benzoxazinorifamycin.”

By “atherosclerosis” is meant the progressive accumulation of smoothmuscle cells, immune cells (e.g., lymphocytes, macrophages, ormonocytes), lipid products (e.g., lipoproteins, or cholesterol),cellular waste products, calcium, or other substances within the innerlining of an artery, resulting in the narrowing or obstruction of theblood vessel and the development of atherosclerosis-associated diseases.Atherosclerosis is typically manifested within large and medium-sizedarteries, and is often characterized by a state of chronic inflammationwithin the arteries.

By “atherosclerosis-associated disease” is meant any disorder that iscaused by or is associated with atherosclerosis. Typically,atherosclerosis of the coronary arteries commonly causes coronary arterydisease, myocardial infarction, coronary thrombosis, and anginapectoris. Atherosclerosis of the arteries supplying the central nervoussystem frequently provokes strokes and transient cerebral ischemia. Inthe peripheral circulation, atherosclerosis causes intermittentclaudication and gangrene and can jeopardize limb viability.Atherosclerosis of an artery of the splanchnic circulation can causemesenteric ischemia. Atherosclerosis can also affect the kidneysdirectly (e.g., renal artery stenosis).

A patient who is being treated for an atherosclerosis-associated diseaseis one who a medical practitioner has diagnosed as having such adisease. Diagnosis may be by any suitable means. Methods for diagnosingatherosclerosis by measuring systemic inflammatory markers aredescribed, for example, in U.S. Pat. No. 6,040,147, hereby incorporatedby reference. Diagnosis and monitoring may employ an electrocardiogram,chest X-ray, echocardiogram, cardiac catheterization, ultrasound (forthe measurement of vessel wall thickness), or measurement of bloodlevels of CPK, CPK-MB, myoglobin, troponin, homocysteine, or C-reactiveprotein. A patient in whom the development of anatherosclerosis-associated disease is being prevented is one who has notreceived such a diagnosis. One in the art will understand that thesepatients may have been subjected to the same tests (electrocardiogram,chest X-ray, etc.) or may have been identified, without examination, asone at high risk due to the presence of one or more risk factors (e.g.,family history, hypertension, diabetes mellitus, high cholesterollevels). Thus, prophylactic administration of a rifamycin analog isconsidered to be preventing the development of anatherosclerosis-associated disease.

An atherosclerosis-associated disease has been treated or prevented whenone or more tests of the disease (e.g., any of the those describedabove) indicate that the patient's condition has improved or thepatient's risk reduced. In one example, a reduction in C-reactiveprotein to normal levels indicates that an atherosclerosis-associateddisease has been treated or prevented.

An alternative means by which treatment or prevention is assessedincludes determination of the presence of an infection of C. pneumoniae.Any suitable method may be employed (e.g., determination of C.pneumoniae in blood monocytes or in the atheroma itself (e.g., inmacrophages or foam cells present in the fatty streak), or detection ofC. pneumoniae DNA, RNA, or antibodies to C. pneumoniae in a biologicalsample from the patient).

By “debris” is meant the mucoid exudate or desquamated epithelium in aninfected ear of a patient having an ear infection.

By “ear wick” is meant a sponge, cotton, gauze, compressedhydroxycellulose, or any other material used to increase the penetrationof rifamycin to the infected otic area. The ear wick is typicallyinserted into the canal under direct vision. Its presence helps wickeardrops along the canal, hold the solution in contact with the skin ofthe canal, and apply pressure to the canal skin.

By “granulation tissue” is meant the highly vascularized tissue thatreplaces the initial fibrin clot in a wound. Vascularization is a resultof an ingrowth of capillary endothelium from the surroundingvasculature. The tissue is also rich in fibroblasts and leucocytes.

“Antibiotic-associated bacterial diarrhea” means the condition whereinantibiotic therapy disturbs the balance of the microbial flora of thegut, allowing pathogenic organisms such as C. difficile to flourish.These organisms cause diarrhea. Antibiotic-associated bacterial diarrheaincludes such conditions as C. difficile associated diarrhea (CDAD) andpseudomembranous colitis.

“Pseudomembranous colitis,” also known as pseudomembranous enterocolitisor enteritis, means the inflammation of the mucous membrane of bothsmall and large intestine with the formation and passage ofpseudomembranous material (composed of fibrin, mucous, necroticepithelial cells and leukocytes) in the stools.

By “autoimmune disease” is meant a disease arising from an immunereaction against self-antigens and directed against the individual's owntissues. Examples of autoimmune diseases include but are not limited tosystemic lupus erythematosus, rheumatoid arthritis, myasthenia gravis,and Graves' disease.

By “bacteria” is meant a unicellular prokaryotic microorganism thatusually multiplies by cell division.

By “bacterial infection” is meant the invasion of a host animal bypathogenic bacteria. For example, the infection may include theexcessive growth of bacteria that are normally present in or on the bodyof an animal or growth of bacteria that are not normally present in oron the animal. More generally, a bacterial infection can be anysituation in which the presence of a bacterial population(s) is damagingto a host animal. Thus, an animal is “suffering” from a bacterialinfection when an excessive amount of a bacterial population is presentin or on the animal's body, or when the presence of a bacterialpopulation(s) is damaging the cells or other tissue of the animal.

By “chronic disease” is meant a disease that is inveterate, of longcontinuance, or progresses slowly, in contrast to an acute disease,which rapidly terminates. A chronic disease may begin with a rapid onsetor in a slow, insidious manner but it tends to persist for severalweeks, months or years, and has a vague and indefinite termination.

By “immunocompromised” is meant a person who exhibits an attenuated orreduced ability to mount a normal cellular or humoral defense tochallenge by infectious agents, e.g., viruses, bacterial, fungi, andprotozoa. Persons considered immunocompromised include malnourishedpatients, patients undergoing surgery and bone narrow transplants,patients undergoing chemotherapy or radiotherapy, neutropenic patients,HIV-infected patients, trauma patients, burn patients, patients withchronic or resistant infections such as those resulting frommyelodysplastic syndrome, and the elderly, all of who may have weakenedimmune systems.

By “inflammatory disease” is meant a disease state characterized by (1)alterations in vascular caliber that lead to an increase in blood flow,(2) structural changes in the microvasculature that permit the plasmaproteins and leukocytes to leave the circulation, and (3) emigration ofthe leukocytes from the microcirculation and their accumulation in thefocus of injury. The classic signs of acute inflammation are erythema,edema, tenderness (hyperalgesia), and pain. Chronic inflammatorydiseases are characterized by infiltration with mononuclear cells (e.g.,macrophages, lymphocytes, and plasma cells), tissue destruction, andfibrosis. Non-limiting examples of inflammatory disease include asthma,coronary artery disease, arthritis, conjunctivitis, lymphogranulomavenerum, and salpingitis.

By “intracytoplasmic inclusion” is meant a replicating reticulate body(RB) that has no cell wall. Such inclusions may be detected, forexample, through chlamydiae sample isolation and propagation onmammalian cell lines, followed by fixing and staining using one of avariety of staining methods including Giemsa staining, iodine staining,and immunofluorescence. These inclusions have a typical round or ovalappearance.

By “persistent bacterial infection” is meant an infection that is notcompletely eradicated through standard treatment regimens usinganti-bacterial agents. Persistent bacterial infections are caused bybacteria capable of establishing a cryptic or latent phase of infectionand may be classified as such by culturing the bacteria from a patientand demonstrating bacterial survival in vitro in the presence ofanti-bacterial agents or by determination of anti-bacterial treatmentfailure in a patient. As used herein, a persistent infection in apatient includes any recurrence of chlamydial infection, after receivinganti-bacterial treatment, from the same species (e.g., C. trachomatis)more than two times over the period of two or more years or thedetection of the cryptic phase of the infection in the patient by themethods described. An in vivo persistent infection can be identifiedthrough the use of a reverse transcriptase polymerase chain reaction(RT-PCR) to demonstrate the presence of 16S rRNA transcripts inbacterially infected cells after treatment with anti-bacterial agents(Antimicrob. Agents Chemother. 12:3288-3297, 2000).

By “replicating phase” is meant the phase of the bacterial cell cyclecharacterized by the presence of an RB. The RB is the activelyreplicating form of the Chlamydia. It contains no cell wall and isdetected as an inclusion in the cell.

As used herein, the term “treating” refers to administering orprescribing a pharmaceutical composition for prophylactic and/ortherapeutic purposes. To “prevent disease” refers to prophylactictreatment of a patient who is not yet ill, but who is susceptible to, orotherwise at risk of, a particular disease. To “treat disease” or usefor “therapeutic treatment” refers to administering treatment to apatient already suffering from a disease to improve the patient'scondition. Thus, in the claims and embodiments, treating is theadministration to an animal either for therapeutic or prophylacticpurposes. An ear infection has been treated when one or more tests ofthe disease (e.g., any of the those described below) indicate that thepatient's condition has improved. The detection of an infection may bedone by a pneumatic otoscopic examination of the patient, or by areduction in infection-associated symptoms in the patient (e.g.,inflammation of ear drums, redness of ear drums, presence of fluid inears). Reduction of symptoms may also be determined, for example, byaudiogram to check recovery from hearing loss. Prophylacticadministration of a rifamycin of the invention is considered to bepreventing the development of an ear infection.

By “effective amount” is meant the amount of a compound required totreat or prevent an infection. The effective amount of activecompound(s) used to practice the present invention for therapeutic orprophylactic treatment of conditions caused by or contributed to by amicrobial infection varies depending upon the manner of administration,the age, body weight, and general health of the subject. Ultimately, theattending physician or veterinarian will decide the appropriate amountand dosage regimen. Such amount is referred to as an “effective” amount.

The term “microbial infection” refers to the invasion of the host animalby pathogenic microbes. This includes the excessive growth of microbesthat are normally present in or on the body of an animal. Moregenerally, a microbial infection can be any situation in which thepresence of a microbial population(s) is damaging to a host animal.Thus, an animal is “suffering” from a microbial infection when excessivenumbers of a microbial population are present in or on an animal's body,or when the presence of a microbial population(s) is damaging the cellsor other tissue of an animal.

The term “microbes” includes, for example, bacteria, fungi, yeasts,viruses and protozoa.

By “intracellular pathogen” is meant an infection by any facultative orobligate intracellular microbe.

By “obligate intracellular pathogen” is meant a microbe that must use anintracellular location (e.g., a host cell) in order to replicate.

By “facultative intracellular pathogen” is meant a microbe that is ableto survive within an intracellular location (e.g., a host cell), butdoes not require an intracellular environment to replicate.

The term “administration” or “administering” refers to a method ofgiving a dosage of a pharmaceutical composition to an animal, where themethod is, e.g., topical, oral, intravenous, intraperitoneal, orintramuscular. The preferred method of administration can vary dependingon various factors, e.g., the components of the pharmaceuticalcomposition, site of the potential or actual disease, and severity ofdisease.

The terms “animal,” “subject,” and “patient” specifically includehumans, cattle, horses, dogs, cats, and birds, but also can include manyother species.

DETAILED DESCRIPTION

We have discovered that rifamycin analogs are useful for treating orpreventing a variety of microbial infections. The compounds of thepresent invention are described by formula (I):

or a pharmaceutically acceptable salt thereof, wherein

-   -   A is H, OH, O—(C₁-C₆ alkyl), or O—(C₁-C₄ alkaryl);    -   W is O, S, or NR¹,wherein R¹ is H or C₁-C₆ alkyl;    -   X is H or COR², wherein R² is C₁-C₆ alkyl, which can be        substituted with from 1 to 5 hydroxyl groups, or O—(C₃-C₇        alkyl), which can be substituted with from 1 to 4 hydroxyl        groups, wherein each alkyl carbon is bound to no more than one        oxygen;    -   Y is H, OR³, or Hal;    -   Z is H, OR³, or Hal;    -   R³ is C₁-C₆ alkyl; and    -   R⁴ has the formula:    -   when each of m and n is 1,    -   each of R⁵ and R⁶ is H, or R⁵ and R⁶ together are ═O;    -   R⁷ and R¹⁰ together form a single bond or a C₁-C₃ linkage, which        optionally contains a non-vicinal O, S, or N(R²³), R⁷ and R¹²        together form a single bond or a C₁-C₂ linkage, which optionally        contains a non-vicinal O, S, or N(R²³), or R⁷ and R¹⁴ together        form a single bond or a C₁ linkage, wherein R²³ is H, C₁-C₆        alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴,        CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂        aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl;    -   R⁸ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R⁸ and R¹² together        form a single bond, or R⁸ and R⁹ together are ═N—O R¹⁸, wherein        R¹⁸ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl;    -   R⁹ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R⁹ and R⁸ together        are ═N—OR¹⁸;    -   R¹⁰ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R¹⁰ and R⁷ together        form a single bond or a C₁-C₃ linkage, which optionally contains        a non-vicinal O, S, or N(R²³), or R¹⁰ and R¹⁶ together form a        C₁-C₂ alkyl linkage, which optionally contains a non-vicinal O,        S, or N(R²³), or R¹⁰ and R¹⁷ together form a C₁-C₃ alkyl        linkage, which optionally contains a non-vicinal O, S, or        N(R²³), or R¹⁰ and R¹¹ together are ═O, wherein R²³ is H, C₁-C₆        alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴,        CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂        aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl;    -   R¹¹ is H, or R¹¹ and R¹⁰ together are ═O;    -   R¹² is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R¹² and R⁷together        form a single bond or a C₁-C₂ linkage, which optionally contains        a non-vicinal O, S, or N(R²³), or R¹² and R⁸ together form a        single bond, or R¹² and R¹⁶ together form a C₂-C₄ alkyl linkage,        which optionally contains a non-vicinal O, S, or N(R²³), wherein        R²³ is H, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴,        CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆        alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄        alkheteroaryl;        -   each of R¹³ and R¹⁵ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl;        -   R¹⁴ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R¹⁴ and R⁷            together form a single bond or a C₁ linkage;        -   R¹⁶ is H, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₆-C₁₂ aryl,            heteroaryl, C₁-C₄ alkaryl, or C₁-C₄ alkheteroaryl, or R¹⁶            and R¹² together form a C₂-C₄ alkyl linkage, which            optionally contains a non-vicinal O, S, or N(R²³), or R¹⁶            and R¹⁰ together form a C₁-C₂ alkyl linkage, which            optionally contains a non-vicinal O, S, or N(R²³), wherein            R²³ is H, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴,            COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is            C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or            C₁-C₄ alkheteroaryl; and        -   R¹⁷ is H, C₁-C₆ alkyl, COR¹⁹, CO₂R¹⁹, or CONHR¹⁹, CSR¹⁹,            COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or SO₂NHR¹⁹, wherein R¹⁹ is            C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or            C₁-C₄ alkheteroaryl, or R¹⁷ and R¹⁰ together from C₁-C₃            alkyl linkage, which optionally contains a non-vicinal O, S,            or N(R²³), wherein R²³ is H, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or            CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or            SO₂NHR²⁴, wherein R²⁴ is C₂-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄            alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl; or        -   when m is 0 and n is 1,        -   R⁷ and R¹⁰ together form a single bond or a C₁-C₄ linkage,            which optionally contains a non-vicinal O, S, or N(R²³), R⁷            and R¹ ² together form a single bond or a C₁-C₃ linkage,            which optionally contains a non-vicinal O, S, or N(R²³), or            R⁷ and R¹⁴ together form a single bond or a C₁-C₂ linkage,            which optionally contains a non-vicinal O, S, or N(R²³),            wherein R²³ is H, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴,            CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein            R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl,            or C₁-C₄ alkheteroaryl;    -   each of R⁸, R⁹, and R¹¹ is H;    -   R¹⁵ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl;    -   R¹⁰ is H or R¹⁰ and R⁷ together form a single bond or a C₁-C₄        linkage, which optionally contains a non-vicinal O, S, or        N(R²³), wherein R²³ is H, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or        CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴,        wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl,        heteroaryl, or C₁-C₄ alkheteroaryl;    -   R¹² is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R¹² and R⁷ together        form a single bond or a C₁-C₃ linkage, which optionally contains        a non-vicinal O, S, or N(R²³), R¹² and R¹ ³ together form a        —CH₂CH₂— linkage, or R¹² and R¹⁶ together form a C₂-C₄ alkyl        linkage, which optionally contains a non-vicinal O, S, or        N(R²³), wherein R²³ is H, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or        CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴,        wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl,        heteroaryl, or C₁-C₄ alkheteroaryl;    -   R¹³ is H, C₁-C₆ alkyl, C₁-C₄ alkaryl, or R¹³ and R¹²together        form a —CH₂CH₂— linkage;    -   R¹⁴ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R¹⁴ and R⁷ together        form a single bond or a C₁-C₂ linkage, which optionally contains        a non-vicinal O, S, or N(R²³), wherein R²³ is H, C₁-C₆ alkyl,        COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴,        SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl,        C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl;    -   R¹⁶ is H, C₁-C₆ alkyl, C₁-C₆ alkoxy, C₆-C₁₂ aryl, heteroaryl,        C₁-C₄ alkaryl, or C₁-C₄ alkheteroaryl, or R¹⁶ and R¹² together        form a C₂-C₄ alkyl linkage, which optionally contains a        non-vicinal O, S, or N(R²³), wherein R²³ is H, C₁-C₆ alkyl,        COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴,        SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl,        C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl; and    -   R¹⁷ is H, C₁-C₆ alkyl, COR¹⁹, CO₂R¹⁹, or CONHR¹⁹, CSR¹⁹, COSR¹⁹,        CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or SO₂NHR¹⁹, wherein R¹⁹ is C₁-C₆        alkyl, C₆-C₁₂ aryl, C₁C₄ alkaryl, heteroaryl, or C₁-C₄        alkheteroaryl; or    -   A is OH;    -   X is H;    -   W, Y, and Z are defined as above; and    -   R⁴ is selected from the group consisting of:        wherein R²¹ is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄        alkaryl, or C₁-C₄ alkheteroaryl, R²⁰ is H, C₁-C₆ alkyl, COR¹⁹,        CO₂R¹⁹, or CONHR¹⁹, CSR¹⁹, COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or        SO₂NHR¹⁹, wherein R¹⁹ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄        alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl; or    -   A is OH;    -   X is COCH₃;    -   W, Y, and Z are defined as above; and    -   R⁴ is selected from the group consisting of:        wherein R²¹ is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄        alkaryl, or C₁-C₄ alkheteroaryl, R²⁰ is H, C₁-C₆ alkyl, COR¹⁹,        CO₂R¹⁹, or CONHR¹⁹, CSR¹⁹, COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or        SO₂NHR¹⁹, wherein R¹⁹ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl        heteroaryl, or C₁-C₄ alkheteroaryl; or    -   A is H or OH;    -   X is H or COCH₃;    -   W, Y, and Z are defined as above; and    -   R⁴ is        with the proviso that one or both of Y and Z are Hal, or    -   A is H or OH;    -   X is H or COCH₃;    -   W, Y, and Z are defined as above; and    -   R⁴ is        wherein R²² is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄        alkaryl, C₁-C₄ alkheteroaryl, COR²⁴, CO₂R²⁴, CONHR²⁴, CSR²⁴,        COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is        C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄        alkheteroaryl, and r is 1-2, or    -   A is H or OH;    -   X is H or COCH₃;    -   W, Y, and Z are defined as above; and    -   R⁴ is        wherein R²¹ is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄        alkaryl, or C₁-C₄ alkheteroaryl, or    -   A is H or OH;    -   X is H or COCH₃;    -   W, Y, and Z are defined as above; and    -   R⁴ is        wherein ═E is ═O or (H,H), R²² is H, C₁-C₆ alkyl, C₆-C₁₂ aryl,        heteroaryl, C₁-C₄ alkaryl, C₁-C₄ alkheteroaryl, COR²⁴, CO₂R²⁴,        CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂ R²⁴, or S₂NHR²⁴,        wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl,        heteroaryl, or C₁-C₄ alkheteroaryl, r is 1-2, and s is 0-1, or    -   A is H or OH;    -   X is H or COCH₃;    -   W, Y, and Z are as described above;    -   and R⁴ is

For those compounds in which R⁴ has the formula:

several different ring systems can be constructed from this genericformula. In one example, compounds having formula (A) are constructedwhen each of m and n is 1 and R⁷ forms a single bond with R¹⁴.

In another example, compounds having formula (B) are constructed wheneach of m and n is 1, R⁷ forms a single bond with R¹⁴, and R⁸ forms asingle bond with R¹².

In another example, compounds having formula (C) are constructed when mis 0 and n is 1, R⁷ forms a single bond with R¹⁴, and R¹² forms a C₃alkyl linkage with R¹⁶.

In another example, compounds having formula (D) are constructed when mis 0, n is 1, and R⁷ forms a single bond with R¹⁴.

In another example, compounds having formula (E) are constructed wheneach of m and n is 1 and R⁷ forms a single bond with R¹².

In another example, compounds having formula (F) are constructed wheneach of m and n is 1, R⁷ forms a single bond with R¹², and R⁸ forms a C₁linkage with R¹⁶.

In yet another example, compounds having formula (G) are constructedwhen m is 0 and n is 1, R⁷ forms a single bond with R¹⁴, and R¹² forms aC₂ alkyl linkage, containing an NR²³ moiety, with R¹⁶.

We have identified a method of preventing, stabilizing, or inhibitingthe growth of microbes, or killing microbes. The method involvescontacting microbes or a site susceptible to microbial growth with acompound of the invention. Compounds of the present invention can beused to treat, stabilize or prevent a microbial infection in an animal.In this method, the step of contacting microbes or a site susceptible tomicrobial infection (e.g., a site in or on the body of an animal) with acompound of the invention includes administering to the animal thecompound in an amount sufficient to treat, stabilize, or prevent themicrobial infection in the animal. In a related aspect, the inventionfeatures a method of treating any disease associated with such amicrobial infection.

Compounds of the present invention can be used to treat atherosclerosisor diseases associated therewith, sexually transmitted diseases caused,for example, by C. trachomatis or N. gonorrhoeae, otitis media and otherear infections, antibiotic-associated colitis, gastritis and ulcersassociated with an infection of H. pylori, Gram-positive infections,community-acquired pneumonia, upper and lower respiratory tractinfections, skin and soft tissue infections, bone and joint infections,hospital-acquired lung infections, urinary tract infections,pyelonephritis, intra-abdominal infections, bacteremia, bacterialsepsis, would infections, peritonitis, osteomyelitis, infections afterbums, pelvic inflammatory disease, and diseases associated with chronicinfections.

Atherosclerosis and other Diseases Associated with Chlamydial Infection

An association was previously reported in International Publication No.WO 98/50074 between the cryptic phase of a persistent chlamydialinfection of body fluids and/or tissues and several chronic diseasesyndromes of previously unknown etiology in humans. To date, thesediseases include, but are not limited to, atherosclerosis, multiplesclerosis, rheumatoid arthritis, inflammatory bowel disease,interstitial cystitis, fibromyalgia, autonomic nervous dysfunction(neural-mediated hypotension); pyoderma gangrenosum, and chronic fatiguesyndrome.

As described in International Publication No. WO 98/50074, several linesof evidence have led to the establishment of a link between Chlamydiaand a broad set of inflammatory, autoimmune, and immune deficiencydiseases. These include (i) the association between the cryptic phase ofa persistent chlamydial infection of body fluids and/or tissues andseveral chronic disease syndromes as described above, (ii) publishedevidence of an association between atherosclerosis and Chlamydia(Circulation, 96:404-407, 1997), and (iii) an understanding of theimpact the persistent infection established by the cryptic phase ofchlamydial infections can have on infected cells and the immune system.Thus, the present invention describes methods for treating chronicdiseases associated with the cryptic phase of a persistent chlamydialinfection, such as autoimmune diseases, inflammatory diseases anddiseases that occur in immunocompromised individuals by treating thecryptic phase of the infection in an individual in need thereof, usingthe rifamycin analogs described herein. Progress of the treatment can beevaluated, using the diagnostic tests described herein, to determine thepresence or absence of Chlamydia. Physical improvement in the conditionsand symptoms typically associated with the disease to be treated canalso be evaluated. Based upon these evaluating factors, the physiciancan maintain or modify the anti-bacterial therapy accordingly.

The therapies described herein can be used for the treatment of chronicimmune and autoimmune diseases when patients are demonstrated to have aChlamydia load by the methods of detection described above. Thesediseases include, but are not limited to, chronic hepatitis, systemiclupus erythematosus, arthritis, thyroidosis, scleroderma, diabetesmellitus, Graves' disease, Beschet's disease, and graft versus hostdisease (graft rejection). The therapies of this invention can also beused to treat any disorders in which a chlamydial species is a factor orco-factor.

Thus, the present invention can be used to treat a range of disorders inaddition to the above immune and autoimmune diseases when demonstratedto be associated with chlamydial infection by the methods of detectiondescribed herein; for example, various infections, many of which produceinflammation as primary or secondary symptoms, including, but notlimited to, sepsis syndrome, cachexia, circulatory collapse and shockresulting from acute or chronic bacterial infection, acute and chronicparasitic and/or infectious diseases from bacterial, viral or fungalsources, such as a HIV, AIDS (including symptoms of cachexia, autoimmunedisorders, AIDS dementia complex and infections) can be treated, as wellas Wegners Granulomatosis.

Among the various inflammatory diseases, there are certain features thatare generally agreed to be characteristic of the inflammatory process.These include fenestration of the microvasculature, leakage of theelements of blood into the interstitial spaces, and migration ofleukocytes into the inflamed tissue. On a macroscopic level, this isusually accompanied by the familiar clinical signs of erythema, edema,tenderness (hyperalgesia), and pain. Inflammatory diseases, such aschronic inflammatory pathologies and vascular inflammatory pathologies,including chronic inflammatory pathologies such as aneurysms,hemorrhoids, sarcoidosis, chronic inflammatory bowel disease, ulcerativecolitis, and Crohn's disease and vascular inflammatory pathologies, suchas, but not limited to, disseminated intravascular coagulation,atherosclerosis, and Kawasaki's pathology are also suitable fortreatment by methods described herein. The invention can also be used totreat inflammatory diseases such as coronary artery disease,hypertension, stroke, asthma, chronic hepatitis, multiple, sclerosis,peripheral neuropathy, chronic or recurrent sore throat, laryngitis,tracheobronchitis, chronic vascular headaches (including migraines,cluster headaches and tension headaches), and pneumonia.

Treatable disorders when associated with a chlamydial infection alsoinclude, but are not limited to, neurodegenerative diseases, including,but not limited to, demyelinating diseases, such as multiple sclerosisand acute transverse myelitis; extrapyramidal and cerebellar disorders,such as lesions of the corticospinal system; disorders of the basalganglia or cerebellar disorders; hyperkinetic movement disorders such asHuntington's Chorea and senile chorea; drug-induced movement disorders,such as those induced by drugs which block CNS dopamine receptors;hypokinetic movement disorders, such as Parkinson's disease; progressivesupranucleo palsy; cerebellar and spinocerebellar disorders, such asastructural lesions of the cerebellum; spinocerebellar degenerations(spinal ataxia, Friedreich's ataxia, cerebellar cortical degenerations,multiple systems degenerations (Mencel, Dejerine-Thomas, Shi-Drager, andMachado Joseph)); and systemic disorders (Refsum's disease,abetalipoprotemia, ataxia, telangiectasia, and mitochondrialmulti-system disorder); demyelinating core disorders, such as multiplesclerosis, acute transverse myelitis; disorders of the motor unit, suchas neurogenic muscular atrophies (anterior horn cell degeneration, suchas amyotrophic lateral sclerosis, infantile spinal muscular atrophy andjuvenile spinal muscular atrophy); Alzheimer's disease; Down's Syndromein middle age; Diffuse Lewy body disease; senile dementia of Lewy bodytype; Wernicke-Korsakoff syndrome; chronic alcoholism; Creutzfeldt-Jakobdisease; subacute sclerosing panencephalitis, Hallerrorden-Spatzdisease; and Dementia pugilistica, or any subset thereof.

It is also recognized that malignant pathologies involving tumors orother malignancies, such as, but not limited to leukemias (acute,chronic myelocytic, chronic lymphocytic and/or myelodyspastic syndrome);lymphomas (Hodgkin's and non-Hodgkin's lymphomas, such as malignantlymphomas (Burkitt's lymphoma or mycosis fungoides)); carcinomas (suchas colon carcinoma) and metastases thereof, cancer-related angiogenesis;infantile hemangiomas; and alcohol-induced hepatitis. Ocularneovascularization, psoriasis, duodenal ulcers, and angiogenesis of thefemale reproductive tract can also be treated when demonstrated by thediagnostic procedures described herein to be associated with achlamydial infection.

Ear Infections

Ear infections typically affect the middle or the external ear andinclude, for example, otitis media, otitis externa, and infectionscaused by surgical interventions. Due to multiplicity of secondarycomplications that arise from ear infections such as hearing loss, thetreatment and prevention of such conditions is critical.

Topical administration of a compound of formula (I) is effective intreating or preventing an infection of the ear, such as otitis media orotitis externa. In the case of otitis media or externa, infections areprimarily caused by H. influenza, M. catarhalis, S. pneumoniae, S.pyogenes, S. intermedius, S. epidermidis, S. aureus, S. caprae, S.auriculis, S. capitis, S. haemolytis, P. aeroginosa, P. mirabilis, P.vulgaris, E. faecalis, or E. coli. A compound of formula (I) can be usedto treat each of these infections of the ear. The rifamycin analog offormula (I) may, for example, be topically administered to the area ofthe ear to which surgical intervention was performed or, alternatively,the rifamycin may be administered to the ear of the patientprophylactically, prior to otic surgery, noninvasive otic procedures, orother types of surgery. Exemplary surgical procedures include forexample, cochlear implant surgery, tympanoplasty, tympanostomy tubeinsertion, removal of tumors (e.g., cholesteatoma), or stapedectomy. Thecompound may be administered to the area of the ear to which surgicalintervention will be performed, for example, within seven days, twodays, one day, 12 hours, 10 hours, 6 hours, 4 hours, 2 hours, 1 hour, orless than 1 hour prior to or following the surgical intervention. Thecompositions may be used for acute treatment of temporary conditions, ormay be administered chronically.

A compound of formula (I) may be given daily (e.g., once, twice, threetimes, or four times daily) or less frequently (e.g., once every otherday, or once or twice weekly). Typically, patients are administered adosage consisting of one to four drops of solution containing thecompound. The compound may be contained in any appropriate amount in anysuitable carrier substance, and is generally present in an amountbetween 0.001% and 5%, desirably 0.01% and 3%, more desirably 0.1% and1%, and even more desirably 0.1% and 0.4% by weight of the total volume(w/v) of the composition. The compound is provided in a dosage form thatis suitable for topical administration. Thus, a composition containing acompound of formula (I) may be in the form of a solution, aerosol, gel,ointment, nebulizer, or suspension. Alternatively, the rifamycin analogmay be administered by placing an impregnated porous media into theexternal ear canal to the tympanic membrane. The pharmaceuticalcomposition can generally be formulated according to conventionalpharmaceutical practice.

Aural Toilet

The external auditory canal and tissues lateral to the infected middleear often are covered with mucoid exudate or desquamated epithelium.Since topically applied preparations cannot generally penetrate affectedtissues until these interposing materials are removed, aural toilet isdesirably performed before administering a compound of formula (I).Aural toilet may be performed by a health provider, the patient, or anyother individual. Removal of debris may be performed mechanically withthe assistance of a microscope and microinstruments. Aural irrigationmay also be performed using a solution containing peroxide. Theconcentration of peroxide should be the highest concentration withoutcausing significant pain, or discomfort, to the patient. As an example,a solution of 50% peroxide and 50% sterile water can be used. Thirty to40 mL of this solution can be irrigated through the external auditorycanal, using a small syringe or bulb-type aspirator. The irrigantsolution is allowed to drain out (e.g., for 5-10 minutes) prior toadministering a rifamycin of the present invention.

Granulation Tissue

Granulation tissue often fills the middle ear and medial portions of theexternal auditory canal, and reducing this accumulation is beneficialfor resolution of an ear infection. Granulation tissue may also preventtopically applied antimicrobial agents from penetrating to the site ofinfection, and the amount of granulation tissue is desirably reducedthroughout the regimen. Although topical antimicrobial drops can reducegranulation by eliminating infection and by removing the incitingirritating inflammation, the amount of granulation tissue may be reducedusing other methods known in the art. For example, topical steroids mayhasten the resolution of middle ear granulation, thus improvingpenetration of topically delivered antibiotics.

Cautery may also be used to reduce the amount of granulation tissue andto reduce its formation. Microbipolar cautery may be administered by ahealth provider. Chemical cautery, using for example silver nitrate, mayalso be applied to an infected ear in the form of silver nitrate sticks.Excision of granulation tissue may also be performed by a health careprovider with a microscope and microinstruments.

Ear Canal Acidification

In a patient affected with otitis externa, a therapy involving ear canalacidification to restore the physiological acidity of the ear may beperformed. The affected ear is administered with a solution containingacetic acid, which may also include a steroid (e.g., hydrocortisone),aluminum acetate, or rubbing alcohol.

Topical Formulations

Pharmaceutical compositions according to the present invention can beformulated for topical administration to the ear of the patient.Patients having an ear infection may be administered with effectiveamounts of a rifamycin analog of the invention, by means of a solution(e.g., drops), ointment, gel, or aerosol (e.g., nebulizer). Thecomposition is typically administered to the affected otic area bytopically applying, for example, one to four drops of a solution orsuspension, or a comparable amount of an ointment, gel, or other solidor semisolid composition, once, twice, three times, or more than threetimes per day. A porous media or an ear wick (e.g., cotton, gauze, orcompressed hydroxycellulose) may also be used to increase thepenetration of the rifamycin to the infected otic area. The ear wick,which is inserted into the canal under direct vision, is typically adried sponge that. helps wick eardrops along the canal, hold thesolution in contact with the skin of the canal and apply pressure to thecanal skin. Wicks may be removed at one day, two days, or more than twodays, and may be replaced if necessary. Alternatively, the ear wick mayitself be impregnated with a rifamycin analog of the invention. Theseformulations can be made according to known and conventional methods forpreparing such formulations.

Since some of the compounds of formula (I) of this invention may not behighly soluble in water at physiological conditions, a solubilizingexcipient may be used to increase solubility. Solubilization is taken tomean an improvement in the solubility by virtue of surface-activecompounds that can convert substances that are insoluble or virtuallyinsoluble in water into clear, or opalescent, aqueous solutions withoutchanging the chemical structure of these substances in the process.Excipients used for this purpose are restricted to those that are safefor administration to humans. Typically such co-solvents are employed ata level of about 0.01% to 2% by weight.

A variety of solubilizing excipients may be used for the formulation ofthe rifamycin analog, including compounds belonging to the followingclasses: polyethoxylated fatty acids, PEG-fatty acid diesters, PEG-fattyacid mono-ester and di-ester mixtures, polyethylene glycol glycerolfatty acid esters, alcohol-oil transesterification products,polyglycerized fatty acids, propylene glycol fatty acid esters, mixturesof propylene glycol esters and glycerol esters, mono- and diglycerides,sterol and sterol derivatives, polyethylene glycol sorbitan fatty acidesters, polyethylene glycol alkyl ethers, sugar esters, polyethyleneglycol alkyl phenols, polyoxyethylene-polyoxypropylene block copolymers,sorbitan fatty acid esters, lower alcohol fatty acid esters, or ionicsurfactants. Such excipients are described for example, in U.S. patentapplication Ser. No: 60/385,532, hereby incorporated by reference.

Ototopical preparations may vary in viscosity. The use of viscosityenhancing agents to provide the compositions of the invention withviscosities greater than the viscosity of simple aqueous solutions maybe desirable to increase the retention time in the ear. Suchviscosity-building agents include, for example, polyvinyl alcohol,polyvinyl pyrrolidone, methyl cellulose, hydroxypropyl methylcellulose,hydroxyethyl cellulose, carboxymethyl cellulose, hydroxypropylcellulose, or other agents known to those skilled in the art. Suchagents are typically employed at a level of about 0.01% to 2% by weight.Optionally, these preparations may include a buffering agent to maintainan acidic pH, since the normal environment of the external auditorycanal is acidic. However, if treatment is required in the middle earwhere the pH is neutral, the pH can be adjusted accordingly.

Otic pharmaceutical products are typically packaged in multidose form.Preservatives are thus desired to prevent microbial contamination duringuse. Suitable preservatives include: polyquaternium-1, benzalkoniumchloride, thimerosal, chlorobutanol, methyl paraben, propyl paraben,phenylethyl alcohol, edetate disodium, sorbic acid, or other agentsknown to those skilled in the art. Typically such preservatives areemployed at a level of from 0.001% to 1.0% by weight.

A penetration enhancer may also be used to facilitate the diffusion of arifamycin analog of the invention through the tympanic membrane into themiddle and inner ear in order to reduce inflammation of ear tissues. Apenetration enhancer is an agent used to increase the permeability ofthe skin to a pharmacologically active agent to increase the rate atwhich the drug diffuses through the skin and enters the tissues andbloodstream. A chemical skin penetration enhancer increases skinpermeability by reversibly damaging or by altering the physiochemicalnature of the stratum corneum to reduce its diffusional resistance(Osborne D W, Henke J J, Pharmaceutical Technology, November 1997, pp58-86). Examples of penetration enhancers include without limitation:alcohols, such as ethanol and isopropanol; polyols, such as n-alkanols,limonene, terpenes, dioxolane, propylene glycol, ethylene glycol, otherglycols, and glycerol; sulfoxides, such as dimethylsulfoxide (DMSO),dimethylformamide, methyl dodecyl sulfoxide, dimethylacetamide; esters,such as isopropyl myristate/palmitate, ethyl acetate, butyl acetate,methyl proprionate, and capric/caprylic triglycerides; ketones; amides,such as acetamides; oleates, such as triolein; various surfactants, suchas sodium lauryl sulfate; various alkanoic acids, such as caprylic acid;lactam compounds, such as azone; alkanols, such as oleyl alcohol;dialkylamino acetates, and admixtures thereof. The use of suchpenetration enhancers is disclosed, for example, in U.S. Pat. No6,093,417, hereby incorporated by reference.

Other Therapeutic Agents

Compositions containing a compound of the invention may also include asecond therapeutic agent, including for example, another rifamycin, ananesthetic, an antimicrobial agent, a zinc salt, or an anti-inflammatoryagent (e.g., an non-steroidal anti-inflammatory or a steroid). Whenadmixing an antimicrobial agent, the antimicrobial agent is preferablypenicillin G, penicillin V, methicillin, oxacillin, cloxacillin,dicloxacillin, nafcillin, ampicillin, amoxicillin, carbenicillin,ticarcillin, mezlocillin, piperacillin, azlocillin, temocillin,cepalothin, cephapirin, cephradine, cephaloridine, cefazolin,cefamandole, cefuroxime, cephalexin, cefprozil, cefaclor, loracarbef,cefoxitin, cefmatozole, cefotaxime, ceftizoxime, ceftriaxone,cefoperazone, ceftazidime, cefixime, cefpodoxime, ceftibuten, cefdinir,cefpirome, cefepime, BAL5788, BAL9141, imipenem, ertapenem, meropenem,astreonam, clavulanate, sulbactam, tazobactam, streptomycin, neomycin,kanamycin, paromycin, gentamicin, tobramycin, amikacin, netilmicin,spectinomycin, sisomicin, dibekalin, isepamicin, tetracycline,chlortetracycline, demeclocycline, minocycline, oxytetracycline,methacycline, doxycycline, erythromycin, azithromycin, clarithromycin,telithromycin, ABT-773, lincomycin, clindamycin, vancomycin,oritavancin, dalbavancin, teicoplanin, quinupristin and dalfopristin,sulphanilamide, para-aminobenzoic acid, sulfadiazine, sulfisoxazole,sulfamethoxazole, sulfathalidine, linezolid, nalidixic acid, oxolinicacid, norfloxacin, perfloxacin, enoxacin, ofloxacin, ciprofloxacin,temafloxacin, lomefloxacin, fleroxacin, grepafloxacin, sparfloxacin,trovafloxacin, clinafloxacin, gatifloxacin, moxifloxacin, gemifloxacin,sitafloxacin, metronidazole, daptomycin, garenoxacin, ramoplanin,faropenem, polymyxin, tigecycline, AZD2563, or trimethoprim. Preferrednon-steroidal anti-inflammatory agents include, for example, detoprofen,diclofenac, diflunisal, etodolac, fenoprofen, flurbiprofen,indomethacin, ketoprofen, mechlofenameate, mefenamic acid, meloxicam,nabumeone, naproxen sodium, oxaprozin, piroxicam, sulindac, tolmeting,celecoxib, rofecoxib, choline salicylate, salsate, sodium salicylate,magnesium salicylate, aspirin, ibuprofen, paracetamol, acetaminophen,and pseudoephedrine, and preferred steroids include, for example,hydrocortisone, prednisone, fluprednisolone, triamcinolone,dexamethasone, betamethasone, cortisone, prednilosone,methylprednisolone, fluocinolone acetonide, flurandrenolone acetonide,and fluorometholone. Preferred anesthetics according to the inventioninclude, for example, benzocaine, butamben picrate, tetracaine,dibucaine, prilocaine, etidocaine, mepivacaine, bupivicaine, andlidocaine. A zinc salt can be zinc sulfate, zinc chloride, zinc acetate,zinc phenol sulfonate, zinc borate, zinc bromide, zinc nitrate, zincglycerophosphate, zinc benzoate, zinc carbonate, zinc citrate, zinchexafluorosilicate, zinc diacetate trihydrate, zinc oxide, zincperoxide, zinc salicylate, zinc silicate, zinc stannate, zinc tannate,zinc titanate, zinc tetrafluoroborate, zinc gluconate, or zincglycinate. All of the therapeutic agents employed in the compositions ofthe present invention can be used in the dose ranges currently known andused for these agents. Different concentrations may be employeddepending on the clinical condition of the patient, the goal of therapy(treatment or prophylaxis), the anticipated duration, and the severityof the infection for which a rifamycin analog of the invention is beingadministered. Additional considerations in dose selection include thetype of infection, age of the patient (e.g., pediatric, adult, orgeriatric), general health, and comorbidity.

Assays

Compounds of the present invention can be screened for antimicrobialactivity by measuring their minimum inhibitory concentration (MIC),using standard MIC in vitro assays (see, for example, Tomioka et al.,Antimicrob. Agents Chemother. 37:67 (1993). Agents can be screenedagainst C. pneumoniae, C. trachomatis, M. tuberculosis (includingmultiple drug resistant strains), M. avium complex, and otherintracellular infectious microbes. Details of a standard MIC assay areprovided in Example 2.

Therapy

The invention features a method of treating or preventing a disease orcondition associated with a microbial infection by administering acompound of formulas (I). Compounds of the present invention may beadministered by any appropriate route for treatment or prevention of adisease or condition associated with a microbial infection,inflammation, or infection derived autoimmune disease. These may beadministered to humans, domestic pets, livestock, or other animals witha pharmaceutically acceptable diluent, carrier, or excipient, in unitdosage form. Administration may be topical, parenteral, intravenous,intra-arterial, subcutaneous, intramuscular, intracranial, intraorbital,ophthalmic, intraventricular, intracapsular, intraspinal,intracisternal, intraperitoneal, intranasal, aerosol, by suppositories,or oral administration.

Therapeutic formulations may be in the form of liquid solutions orsuspensions; for oral administration, formulations may be in the form oftablets or capsules; and for intranasal formulations, in the form ofpowders, nasal drops, or aerosols.

Methods well known in the art for making formulations are found, forexample, in Remington: The Science and Practice of Pharmacy (20th ed.,ed. A.R. Gennaro AR.), Lippincott Williams & Wilkins, 2000. Formulationsfor parenteral administration may, for example, contain excipients,sterile water, or saline, polyalkylene glycols such as polyethyleneglycol, oils of vegetable origin, or hydrogenated napthalenes.Biocompatible, biodegradable lactide polymer, lactide/glycolidecopolymer, or polyoxyethylene-polyoxypropylene copolymers may be used tocontrol the release of the compounds. Nanoparticulate formulations(e.g., biodegradable nanoparticles, solid lipid nanoparticles,liposomes) may be used to control the biodistribution of the compounds.Other potentially useful parenteral delivery systems includeethylene-vinyl acetate copolymer particles, osmotic pumps, implantableinfusion systems, and liposomes. Formulations for inhalation may containexcipients, for example, lactose, or may be aqueous solutionscontaining, for example, polyoxyethylene-9-lauryl ether, glycholate anddeoxycholate, or may be oily solutions for administration in the form ofnasal drops, or as a gel. The concentration of the compound in theformulation will vary depending upon a number of factors, including thedosage of the drug to be administered, and the route of administration.

Formulations for oral use include tablets containing the activeingredient(s) in a mixture with non-toxic pharmaceutically acceptableexcipients. These excipients may be, for example, inert diluents orfillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystallinecellulose, starches including potato starch, calcium carbonate, sodiumchloride, lactose, calcium phosphate, calcium sulfate, or sodiumphosphate); granulating and disintegrating agents (e.g., cellulosederivatives including microcrystalline cellulose, starches includingpotato starch, croscarmellose sodium, alginates, or alginic acid);binding agents (e.g., sucrose, glucose, sorbitol, acacia, alginic acid,sodium alginate, gelatin, starch, pregelatinized starch,microcrystalline cellulose, magnesium aluminum silicate,carboxymethylcellulose sodium, methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethyleneglycol); and lubricating agents, glidants, and antiadhesives (e.g.,magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenatedvegetable oils, or talc). Other pharmaceutically acceptable excipientscan be colorants, flavoring agents, plasticizers, humectants, bufferingagents, and the like.

The tablets may be uncoated or they may be coated by known techniques,optionally to delay disintegration and absorption in thegastrointestinal tract and thereby providing a sustained action over alonger period. The coating may be adapted to release the active drugsubstance in a predetermined pattern (e.g., in order to achieve acontrolled release formulation) or it may be adapted not to release theactive drug substance until after passage of the stomach; this may beachieved by means of an enteric coating (e.g., a pH-sensitive entericpolymer). Desirably, a substantial amount of the drug is released in thelower gastrointestinal tract. The coating may be a sugar coating, a filmcoating (e.g., based on hydroxypropyl methylcellulose, methylcellulose,methyl hydroxyethylcellulose, hydroxypropylcellulose,carboxymethylcellulose, acrylate copolymers, polyethylene glycols and/orpolyvinylpyrrolidone), or a coating based on methacrylic acid copolymer,cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate,hydroxypropyl methylcellulose acetate succinate, polyvinyl acetatephthalate, shellac, and/or ethylcellulose. Furthermore, a time delaymaterial such as, for example, glyceryl monostearate or glyceryldistearate, may be employed.

The solid tablet compositions may include a coating adapted to protectthe composition from unwanted chemical changes (e.g., chemicaldegradation prior to the release of the active drug substance). Thecoating may be applied on the solid dosage form in a similar manner asthat described in Encyclopedia of Pharmaceutical Technology, 20^(th)Edition, Eds. Swarbrick and Boyland, 2000.

Formulations for oral use may also be presented as chewable tablets, oras hard gelatin capsules wherein the active ingredient is mixed with aninert solid diluent (e.g., potato starch, lactose, microcrystallinecellulose, calcium carbonate, calcium phosphate or kaolin), or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example, peanut oil, liquid paraffin, or olive oil.Powders and granulates may be prepared using the ingredients mentionedabove under tablets and capsules in a conventional manner using, e.g., amixer, a fluid bed apparatus or a spray drying equipment.

Controlled release compositions for oral use may be constructed torelease the active drug by controlling the dissolution and/or thediffusion of the active drug substance.

Any of a number of strategies can be pursued in order to obtaincontrolled release in which the rate of release outweighs the rate ofmetabolism of the compound in question. In one example, controlledrelease is obtained by appropriate selection of various formulationparameters and ingredients, including, e.g., various types of controlledrelease compositions and coatings. Thus, the drug is formulated withappropriate excipients into a pharmaceutical composition that, uponadministration, releases the drug in a controlled manner. Examplesinclude single or multiple unit tablet or capsule compositions, oilsolutions, suspensions, emulsions, microcapsules, microspheres,nanoparticles, patches, and liposomes.

Dissolution or diffusion controlled release can be achieved byappropriate coating of a tablet, capsule, pellet, or granulateformulation of compounds, or by incorporating the compound into anappropriate matrix. A controlled release coating may include one or moreof the coating substances mentioned above and/or, e.g., shellac,beeswax, glycowax, castor wax, carnauba wax, stearyl alcohol, glycerylmonostearate, glyceryl distearate, glycerol palmitostearate,ethylcellulose, acrylic resins, dl-polylactic acid, cellulose acetatebutyrate, polyvinyl chloride, polyvinyl acetate, vinyl pyrrolidone,polyethylene, polymethacrylate, methylmethacrylate,2-hydroxymethacrylate, methacrylate hydrogels, 1,3 butylene glycol,ethylene glycol methacrylate, and/or polyethylene glycols. In acontrolled release matrix formulation, the matrix material may alsoinclude, e.g., hydrated methylcellulose, camauba wax and stearylalcohol, carbopol 934, silicone, glyceryl tristearate, methylacrylate-methyl methacrylate, polyvinyl chloride, polyethylene, and/orhalogenated fluorocarbon.

A controlled release composition containing a compound of the inventionmay also be in the form of a buoyant tablet or capsule (i.e., a tabletor capsule that, upon oral administration, floats on top of the gastriccontent for a certain period of time). A buoyant tablet formulation ofthe compound(s) can be prepared by granulating a mixture of the drug(s)with excipients and 20-75% w/w of hydrocolloids, such ashydroxyethylcellulose, hydroxypropylcellulose, orhydroxypropylmethylcellulose. The obtained granules can then becompressed into tablets. On contact with the gastric juice, the tabletforms a substantially water-impermeable gel barrier around its surface.This gel barrier takes part in maintaining a density of less than one,thereby allowing the tablet to remain buoyant in the gastric juice.

Formulations for Colonic Release

Due to the lack of digestive enzymes, colon is considered a suitablesite for the absorption of various drugs. Furthermore, colonic drugdelivery of a compound of the invention allows local, direct treatmentof colonic disease, e.g., ulcerative colitis, Crohn's disease, or coloncancer. In addition, as the colon has a longer retention time, drugabsorption is prolonged, and total bioavailability is increased. SeeSintov et al., Int. J. Pharma. 143:101-106 (1996).

One approach for delivering a compound of the invention to the colon isbased on the different pH of each compartment of the gastrointestinaltract, with the pH of the proximal GI tract being lower than that of thedistal GI tract. Thus, polymers that are insoluble at low pH and solubleat higher pH have been used to deliver drugs to the distal GI tract.U.S. Pat. Nos. 5,401,512 and 5,541,170, and WO 95/11024 describe drugcompositions for selectively releasing the drug in the colon by way ofexploiting the difference in pH between the colon and other parts of theGI tract.

Another approach is based on the fact that transit time through thestomach is approximately 2 hours, whereas transit time through the smallintestine is approximately 4-6 hours. Thus, in this approach, thedelivery system is designed to withhold the release of a compound of theinvention for about 6-8 hours from the time of administration. Forexample, U.S. Pat. Nos. 5,482,718; 4,627,851; 4,693,895; 4,705,515;5,171,580; 5,536,507; and 4,904,474; and EP 621 032 A1, JP 34929/1991A,EP 453 001 A1, and EP 572 942 A2 disclose time dependent drug deliverysystem. They are designed to prevent drug release for a period of timeexpected to be sufficient for the composition to pass through the uppergastrointestinal tract.

In addition, it is well known that enzymes capable of reducing azo bondsor hydrolyzing glycosidic bonds, which are not degraded in the stomachand small intestine, are present in the colon. Thus, another approachfor the colonic delivery of a compound of the invention is to use azobond-containing polymers (azo polymers) or glycosidic bond-containingmaterials.

The glycosidic bond-containing polymers include disaccharides,oligosaccharides, and polysaccharides. U.S. Pat. No. 5,505,966 disclosesa pharmaceutical composition containing calcium pectinate as a majorcomponent and a filler such as pectin, dextran, avicel, or mixturethereof. U.S. Pat. No. 5,525,634 discloses a pharmaceutical compositioncontaining a synthetic or natural polymer that is degradable by acolonic enzyme, herein calcium pectinate is disclosed as an example of anatural polymer. In U.S. Pat. No. 5,505,966, the calcium pectinatecomposition is used in the form of a coacervate pellet. It is believedthat calcium pectinate, which is insoluble in water, is converted to awater-soluble matrix by sodium ions or potassium ions present in thedigestion solution of upper GI. U.S. Pat. No. 5,525,634 describes acompressed tablet formulation that is prepared by pulverizing andcompressing a pharmaceutical composition containing a drug and calciumpectinate. In the composition, the strength of compression largelyaffects the system disintegration through the GI tract. In anotherexample useful for the formulation of compounds of the invention forcolonic release, Adkin et al., Pharm. Res. 14:103-107,1997, describe theaddition of Guar gum or pectin as a binder of calcium pectinatecompressed tablets and coating them with enteric material. In thisexample, Guar gum or pectin is used as a binder for preventing easydisintegration in upper GI to achieve sustained release effect in colon.In other examples useful for the colonic delivery of the compounds ofthe invention, U.S. Pat. No. 4,432,966 discloses a compositioncomprising microcrystalline cellulose and ethyl cellulose; EP 627 173 A1describes a cellulose composition; WO 95/35100 discloses a starchcapsule and a composition comprising an enteric coating; U.S. Pat. No.5,422,121 discloses a composition comprising a guar gum or locust beangum blended with a film forming material, and U.S. Pat. No. 6,413,494describes compositions that contain two polysaccharides, such as, forexample, galactomannan and pectin, which are degradable by colonicenzymes and are useful for colonic release formulations.

The compound may be optionally administered as a pharmaceuticallyacceptable salt, such as a non-toxic acid addition salts or metalcomplexes that are commonly used in the pharmaceutical industry.Examples of acid addition salts include organic acids such as acetic,lactic, pamoic, maleic, citric, malic, ascorbic, succinic, benzoic,palmitic, suberic, salicylic, tartaric, methanesulfonic,toluenesulfonic, or trifluoroacetic acids or the like; polymeric acidssuch as tannic acid, carboxymethyl cellulose, or the like; and inorganicacid such as hydrochloric acid, hydrobromic acid, sulfuric acidphosphoric acid, or the like. Metal complexes include zinc, iron, andthe like.

Pharmaceutical formulations of compounds of the invention describedherein include isomers such as diastereomers and enantiomers, mixturesof isomers, including racemic mixtures, salts, solvates, and polymorphsthereof.

Compounds of the invention may be used in combination with anotherantifungal, antiviral, antibacterial, or antiprotozoan compound orcombinations thereof. The other agent may be employed as a supplementalantimicrobial for the purpose of broadening the activity spectrum of thetherapeutics or to obtain particular effects, such as, to reduce thedevelopment of drug resistant microbes. Compounds of the invention canbe used either alone or in conjunction with other pharmaceuticalcompounds to effectively combat a single infection. For example,compounds of the invention may be used either alone or combined withacyclovir in a combination therapy to treat HSV-1. Compounds of theinvention may also be used either alone or in conjunction with otherpharmaceutical compounds to combat multiple infections. For example,compounds of the invention may be used in combination with one or moreanti-mycobacterial agents agents such as isoniazid, pyrazinamide, orethambutol to treat or prevent intracellular bacterial infections.Compounds of the invention may also be used in combination with Intron Aand/or a biflavanoid for treating Hepatitis B; with gancyclovir,progancyclovir, famcyclovir, foscarnet, vidarabine, cidovir, and/oracyclovir for treating herpes viruses; and with ribavarin, amantidine,and/or rimantidine for treating respiratory viruses.

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how themethods and compounds claimed herein are performed, made, and evaluated,and are intended to be purely exemplary of the invention and are notintended to limit the scope of what the inventors regard as theirinvention.

Syntheses of Benzoxazinorifamycin Compounds

Rifamycin analogs of formula (I) can be synthesized by methods analogousto those disclosed in U.S. Pat. Nos. 4,610,919; 4,983,602; 5,786,349;5,981,522; and 4,859,661, and Chem. Pharm. Bull., 41:148 (1993), each ofwhich is hereby incorporated by reference. Syntheses of rifamycinanalogs of formula (I) are provided in the Examples. The synthesis ofthe 25-hydroxy prodrug analog (see Seligson, et al., Anti-Cancer Drugs12:305-13, 2001),25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-fluoro-5′-[4-isobutyl-1-piperazinyl]benzoxazinorifamycin(Example 5 and FIG. 1) can be used as a guide for the synthesis of otherrifamycin analogs in which the 25-hydroxyl group is derivatized (e.g.,as an ester, a carbamate, or a carbonate).

Benzoxazinorifamycin compounds can be prepared using methods thatrequire the selective protection and deprotection of alcohols, amines,sulfhydryls and/or carboxylic acid functional groups. For example,commonly used protecting groups for amines include carbamates, such astert-butyl, benzyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl,9-fluorenylmethyl, allyl, and m-nitrophenyl. Other commonly usedprotecting groups for amines include amides, such as formamides,acetamides, trifluoroacetamides, sulfonamides, trifluoromethanesulfonylamides, trimethylsilylethanesulfonamides, and tert-butylsulfonyl amides.Examples of commonly used protecting groups for carboxylic acids includeesters, such as methyl, ethyl, tert-butyl, 9-fluorenylmethyl,2-(trimethylsilyl)ethoxy methyl, benzyl, diphenylmethyl, O-nitrobenzyl,ortho-esters, and halo-esters. Examples of commonly used protectinggroups for alcohols include ethers, such as methyl, methoxymethyl,methoxyethoxymethyl, methylthiomethyl, benzyloxymethyl,tetrahydropyranyl, ethoxyethyl, benzyl, 2-napthylmethyl, O-nitrobenzyl,P-nitrobenzyl, P-methoxybenzyl, 9-phenylxanthyl, trityl (includingmethoxy-trityls), and silyl ethers. Examples of commonly used protectinggroups for sulfhydryls include many of the same protecting groups usedfor hydroxyls. In addition, sulfhydryls can be protected in a reducedform (e.g., as disulfides) or an oxidized form (e.g., as sulfonic acids,sulfonic esters, or sulfonic amides). Protecting groups can be chosensuch that selective conditions (e.g., acidic conditions, basicconditions, catalysis by a nucleophile, catalysis by a Lewis acid, orhydrogenation) are required to remove each, exclusive of otherprotecting groups in a molecule. The conditions required for theaddition of protecting groups to amine, alcohol, sulfhydryl, andcarboxylic acid functionalities and the conditions required for theirremoval are provided in detail in T. W. Green and P. G. M. Wuts,Protective Groups in Organic Synthesis (2^(nd) Ed.), John Wiley & Sons,1991 and P. J. Kocienski, Protecting Groups, Georg Thieme Verlag, 1994(hereby incorporated by reference). In the examples that follow, the useof protecting groups is indicated in a structure by the letter P, whereP for any amine, carboxylic acid, sulfhydryl, or alcohol may be any ofthe protecting groups listed above.

EXAMPLE 1

General Coupling Procedure

The synthesis of 5′-substituted benzoxazinorifamycin, 5′-substitutedbenzthiazinorifamycin, and 5′-substituted benzdiazinorifamycin analogscan all proceed through the same general route as shown in Scheme 1,using the general methods disclosed in U.S. Pat. No. 4,965,261 for theattachment of amines to the 5′-position. In this scheme, rifamycinazaquinones of the formula II are dissolved in a suitable solvent, forexample, DMSO, and reacted with amines or the formula III in thepresence of manganese dioxide for several hours at room temperature toform azaquinones of the formula IV. If required, azaquinones of theformula IV can be further reacted with deprotection reagents to remove Xat the 25-position, P′ at the 21 and 23 positions, and/or any P″protecting group introduced with amines of the formula III. In someembodiments, the 25-position can be further derivatized with groups thatintroduce useful pharmacodynamic properties, such as groups thattransform a rifamycin analog into a prodrug. Such groups are known tothose skilled in the art, examples of which can be found in Testa andMayer, Hydrolysis in Drug and Prodrug Metabolism: Chemistry,Biochemistry and Enzymology, published by Vch. Verlagsgesellschaft Mbh.(2003), which is hereby incorporated by reference.

Compound 100 (1.00 g, 1.09 mmol) was dissolved in methyl sulfoxide (10mL) and treated with an amine of the formula V (2.18 mmol) andmanganese(IV) oxide (0.95g, 10.9 mmol) for between 12 h to 120 h at rtto 65° C. The reaction mixture was subsequently diluted with ethylacetate, filtered through celite, washed with water (×3), and dried overNa₂SO₄. Filtration, followed by removal of the solvent in vacuo, yieldeda residue that was purified via flash chromatography (silica gel) usingan appropriate solvent system, such as 1-10% methanol in methylenechloride, to produce a benzoxazinorifamycin of the formula VI. Iffurther purification is desired, a second column employing C₁₈ silicagel, using an appropriate solvent system such as 10% water in methanol,was performed.

Alternatively, using the methods described in Helv. Chim. Acta 56:2369(1973), rifamycin quinones of the formula (VII) can be reacted withanilines of the formula VIII, as shown in Scheme 2, to produceazaquinones of the formula IX.

EXAMPLE 2

General Deacetylation Procedure

Compounds of the formula VI (˜100 mmol) were dissolved in methanol (5mL) and then treated with saturated sodium hydroxide in methanolsolution (5 mL) for between 0.5 h to 3 h at rt. The reaction mixture wasthen poured into saturated ammonium chloride solution and extracted withchloroform. The organics were washed with water (2×) and dried overNa₂SO₄. Filtration, followed by removal of the solvent in vacuo, yieldedthe desired des-acetyl product of the formula X. If desired, theseproducts were purified via flash chromatography (silica gel) using anappropriate solvent system, such as 1-10% methanol in methylenechloride.

EXAMPLE 3

Synthesis of Compound No. 1 (see Table 1 for Structure)

The title compound was prepared using the general coupling procedure ofExample 1 with compound 100 (1.34 g, 1.46 mmol), commercially availablepiperidin-4-yl-carbamic acid, ethyl ester (628 mg, 3.65 mmol), andmanganese(IV) oxide (1.27 g, 14.6 mmol), to provide compound 1 (1.01 g,71% yield) as a blue solid, Mp=218-222° C.; ESI (+) MS: 971 (M+H⁺);UV/Vis: λ_(max)=643.0 nm.

EXAMPLE 4

Synthesis of Compound No. 3 (see Table 1 for Structure)

The title compound was prepared by general deacetylation procedure ofExample 2, using compound 1 (155 mg, 0.160 mmol) to provide compound 3(133 mg, 89% yield) as a blue solid, Mp=206-216° C.; ESI (+) MS: 929(M+H⁺); UV/Vis: λ_(max)=643.0 nm.

EXAMPLE 5

Synthesis of Compound No. 80

The title compound was prepared using the general coupling procedure ofExample 1 with compound 100 (933 mg, 1.02 mmol), commercially available(1S, 4S)-(+)-2,5-diaza-bicyclo[2.2.1]heptane dihydrobromide (531 mg,2.04 mmol), diisopropylethylamine (1.07 mL, 6.12 mmol), andmanganese(IV) oxide (887 mg, 10.2 mmol) to provide compound 80 (206 mg,23% yield) as a blue solid, Mp=>330° C.; ESI (+) MS: 897 (M+H⁺); UV/Vis:λ_(max)=649.4 nm.

EXAMPLE 6

Synthesis of Compound No. 2 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 2 was preparedas follows:

2-(Trimethylsilyl)ethyl p-nitrophenyl carbonate (2.26 g, 7.97 mmol) inabs. ethanol (10 mL) was added to a stirred suspension of compound 101b(1.50 g, 7.97 mmol, J. Chem. Soc., Perkin Trans. 1 2000: 1615-22) in aq.Na₂CO₃ (20 mL, 2M). Water (10 mL) was added and the sides of the flaskwere rinsed with abs. ethanol (15 mL). The reaction was refluxed withstirring for 1 hour, cooled, and then stirred under an atmosphere of N₂at ambient temperature for 16.5 hours. Most of the solvent was removedin vacuo and the resulting slurry was partitioned between methylenechloride and water. The aqueous layer was extracted with methylenechloride (×3). The organic layers were combined and dried over Na₂SO₄.Filtration followed by removal of the solvent in vacuo yielded a residuethat was purified via flash chromatography (19:1, methylenechloride:methanol) to yield compound 101b (2.12 g, 80% yield) as a lightbrown oil, ¹H NMR (CDCl₃, 300 MHz): 0.03 (s, 9H), 0.97 (t,j=8.3 Hz, 2H),1.50 (s, 2H), 2.38 (d,j=8.9 Hz, 2H), 2.92 (s, 1H), 3.06 (d,j=5.9 Hz,2H), 3.55 (s, 2H), 4.14 (t,j=8.2 Hz, 2H), 4.64 (bs, 1H), 7.19-7.30 (m,5H); ESI(+) MS:333(M+H⁺).

A mixture of compound 101b (1.10 g, 3.31 mmol), Pd/C (550 mg, 10%, wet),and methanol (30 mL) was hydrogenated on a Parr apparatus at 50 psi H₂for 20.5 hours. The reaction was filtered through celite, and thesolvent removed in vacuo to yield compound 101 (799 mg, 100% yield) as aviscous yellow oil, ¹H NMR (CDCl₃, 300 MHz): 0.04(s, 9H), 0.98 (t,j=8.4Hz, 2H), 1.63 (s, 2H), 2.39 (d,j=1.8 Hz, 1H), 2.70 (bs, 1H), 2.98(d,j=11.4 Hz, 2H), 3.19 (d,j=11.5 Hz, 2H), 4.15 (t,j=80 Hz, 2H), 5.30(bs, 1H); ESI (+) MS: 243 (M+H⁺).

The title compound was prepared using the general coupling procedure ofExample 1 with compound 100 (1.38 g, 1.51 mmol), amine 101 (730 mg, 3.01mmol), and manganese(IV) oxide (1.31 g, 15.1 mmol), to provide compound2 (830 mg, 53% yield) as a blue solid, Mp=230-231° C.; ESI (+) MS: 1042(M+H⁺); UV/Vis: λ_(max)=640.0 nm.

EXAMPLE 7

Synthesis of Compound No. 81

The title compound was prepared using the general coupling procedure ofExample 1 using compound 100 (1.27 g, 1.39 mmol),2,5-diaza-bicyclo[2.2.2]octane dihydrochloride (515 mg, 2.78 mmol, J.Heterocyclic Chem. 1974, 449-451 and J. Med. Chem. 1974, 481-487),diisopropylethylamine (1.45 mL, 8.34 mmol), and manganese(IV) oxide(1.21 g, 13.9 mmol) to provide compound 81 (54 mg, 4% yield) as a bluesolid, Mp=>300° C.; ESI (+) MS: 911 (M+H⁺); UV/Vis: λ_(max)=653.2 nm.

EXAMPLE 8

Synthesis of Compound No. 4 (see Table 1 for Structure)

A mixture of compound 2 (796 mg, 0.764 mmol), tetrabutylammoniumfluoride (7.64 mL, 1.0 M in THF, 7.64 mmol), and acetonitrile (40 mL)was stirred under an atmosphere of N₂ at 50° C. for 18 hours. Thereaction was cooled and poured over water. The aqueous layer wasextracted with chloroform (×6). The combined organics were washed withwater (×1), then dried over Na₂SO₄. Filtration, followed by removal ofthe solvent in vacuo, yielded a residue that was purified via MPLC(gradient, 1.25-20% methanol:methylene chloride), followed by reversephase chromatography (C₁₈ silica, 9:1, methanol:water) to yield compound4 as a blue solid (521 mg, 76% yield), Mp=216-217° C.; ESI (+) MS: 897(M+H⁺); UV/Vis: λ_(max)=642.3 nm.

EXAMPLE 9

Synthesis of Compound No. 20 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 4 (366 mg, 0.408 mmol) to provide compound20 (232 mg, 66% yield) as a blue solid, Mp=211-231° C.; ESI (+) MS: 855(M+H⁺); UV/Vis: λ_(max)=642.8 nm.

EXAMPLE 10

Synthesis of Compound No. 5 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 5 was preparedas follows:

2-(Trimethylsilyl)ethyl p-nitrophenyl carbonate (2.53 g, 8.92 mmol) inabs. ethanol (25 mL) was added to a stirred suspension of compound 102b(U.S. Pat. No. 5,654,318, 1.93 g, 8.92 mmol) in aq. Na₂CO₃ (20 mL, 2M),followed by the addition of water (10 mL). The reaction was refluxedwith stirring for 1 hour, cooled, and then stirred under an atmosphereof N₂ at ambient temperature for 19 hours. Most of the solvent wasremoved in vacuo and the resulting slurry was partitioned betweenmethylene chloride and water. The aqueous layer was extracted withmethylene chloride (×2). The organic layers were combined and dried overNa₂SO₄. Filtration followed by removal of the solvent in vacuo yielded aresidue that was purified via flash chromatography (19:1, methylenechloride:methanol) to yield compound 102a (2.59 g, 80% yield) as a lightbrown oil, ¹H NMR (CDCl₃, 300 MHz): 0.03 (s, 9H), 0.95-1.01 (m, 2H),1.38-1.76 (m, 5H), 2.13-2.15 (m, 1H), 2.50 (d,j=8.9H), 2.61-2.67 (m,1H), 2.73-2.87 (m, 3H), 3.66 (dd,j=21.5, 13.1 Hz, 2H), 3.93 (bs, 1H),4.12-4.17 (m, 2H), 4.62 (bs, 1H), 7.22-7.31 (m, 5H); ESI (+) MS: 361(M+H⁺).

A mixture of compound 102a (2.57 g, 7.12 mmol), Pd/C (1.3 g, 10%, wet),and methanol (50 mL, sparged with H₂) was stirred under balloon pressureH₂ for 18 hours. The reaction was filtered through celite, and thesolvent removed in vacuo. The resulting residue was purified via MPLC(4:1, methylene chloride:methanol) to yield compound 102 (951 mg, 49%yield) as a yellow oil, ¹H NMR (CDCl₃, 300 MHz): 0.04 (s, 9H), 0.98-1.03(m, 2H), 1.36-1.49 (m, 2H), 1.67-1.72 (m, 2H), 1.71 (bs, 1H), 2.00-2.10(m, 1H), 2.73-2.83 (m, 3H), 3.06-3.18 (m, 2H), 4.00 (d,j=12.3 Hz, 1H),4.15-4.21 (m, 2H), 4.49-4.54 (m, 1H); ESI (+) MS: 271 (M+H⁺).

Teoc-protected precursors to compounds 5 and 12 were prepared by generalcoupling procedure 1 using compound 100 (1.56 g, 1.71 mmol), amine 102(922 mg, 3.41 mmol), and manganese(IV) oxide (1.49 g, 17.1 mmol) toprovide 511 mg (28%) of a blue solid (1:1.1, diastereomer A:diastereomer B) and 601 mg (33%) of a blue solid (diastereomer B).Diastereomer A eluted first using a silica column on MPLC (50-100% ethylacetate:hexanes).

A mixture of the Teoc-protected diastereomer A and B precursors tocompound 5 (1:1.1, diastereomer A: diastereomer B, 363 mg, 0.339 mmol),tetrabutylammonium fluoride (3.40 mL, 1.0 M in THF, 3.39 mmol), andacetonitrile (10 mL) was stirred in a sealed flask at 50° C. for 23hours. The reaction was cooled, and poured over water. The aqueous layerwas extracted with chloroform (×2). The combined organics were washedwith water (×3), then dried over Na₂SO₄. Filtration followed by removalof the solvent in vacuo yielded a residue that was purified via MPLC(gradient, 1.25-5% methanol:methylene chloride) followed by preparatorythin layer chromatography (1.0 mm silica, 9:1, methylenechloride:methanol, ˜50 mg per plate) to yield compound 5 as a blue solid(155 mg), Mp=>300° C.; ESI (+) MS: 925 (M+H⁺); UV/Vis: λ_(max)=650.3 nm.

EXAMPLE 11

Synthesis of Compound No. 12 (see Table 1 for Structure)

A mixture of the Teoc-protected diastereomer B precursor to compound 12from Example 10 (337 mg, 0.315 mmol), tetrabutylammonium fluoride (3.15mL, 1.0 M in THF, 3.15 mmol), and acetonitrile (10 mL) was stirred in asealed flask at 50° C. for 23 hours. The reaction was cooled, and pouredover water. The aqueous layer was extracted with chloroform (×2). Thecombined organics were washed with water (×3), then dried over Na₂SO₄.Filtration followed by removal of the solvent in vacuo yielded a residuethat was purified via MPLC (gradient, 1.25-10% methanol:methylenechloride) followed by preparatory thin layer chromatography (1.0 mmsilica, 9:1, methylene chloride:methanol, ˜50 mg per plate) to yieldcompound 12 as a blue solid (162 mg), Mp=>320° C.; ESI (+) MS: 925(M+H⁺); UV/Vis: λ_(max)=650.5 nm.

EXAMPLE 12

Synthesis of Compound No. 7 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 5 (66 mg, 0.0713 mmol) to provide compound 7(62 mg, 98% yield) as a blue solid, Mp=240-243° C.; ESI (+) MS: 883(M+H⁺); UV/Vis: λ_(max)=650.2 nm.

EXAMPLE 13

Synthesis of Compound No. 15 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 12 (103 mg, 0.111 mmol) to provide 98 mg ofcompound 15 (100% yield) as a blue solid, Mp=216-227° C.; ESI (+) MS:883 (M+H⁺); UV/Vis: λ_(max)=650.3 nm.

EXAMPLE 14

Synthesis of Compound No. 6 (see Table 1 for Structure)

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (2.00 g, 2.18 mmol), commercially availableN-phenyl-N-(4-piperidinyl)propanamide (1.01 g, 4.36 mmol), andmanganese(IV) oxide (1.93 g, 22.1 mmol) to provide compound 6 (1.35 g,60% yield) as a blue solid, Mp=224-228° C.; ESI (+) MS: 1031 (M+H⁺);UV/Vis: λ_(max)=642.5 nm.

EXAMPLE 15

Synthesis of Compound No. 8 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 6 (0.320 g, 0.310 mmol) to provide compound8 (0.301 g, 98% yield) as a blue solid, Mp=214-216° C.; ESI (+) MS: 957(M+H⁺); UV/Vis: λ_(max)=642.8 nm.

EXAMPLE 16

Synthesis of Compound No. 9 (see Table 1 for Structure)

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (2.50 g, 2.73 mmol), commercially available4-(piperidin-4-yl)-morpholine (1.00 g, 5.87 mmol), and manganese(IV)oxide (2.40 g, 27.6 mmol) to provide compound 9 (1.26 g, 47% yield) as ablue solid, Mp=228-230° C.; ESI (+) MS: 969 (M+H⁺); UV/Vis:λ_(max)=646.0 nm.

EXAMPLE 17

Synthesis of Compound No. 11 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 9 (0.410 g, 0.420 mmol) to provide compound11 (0.380 g, 96% yield) as a blue solid, Mp=222-224° C.;, ESI (+) MS:927 (M+H⁺); UV/Vis: λ_(max)=646.2 nm.

EXAMPLE 18

Synthesis of Compound No. 10 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 10 was preparedas follows:

2-(Trimethylsilyl)ethyl p-nitrophenyl carbonate (1.35 g, 4.78 mmol) inacetonitrile (10 mL) was added to a stirred solution of compound 104b(Tet. Lett. 2002, 899-902, 920 mg, 4.55 mmol), and diisopropylethylamine(0.833 mL, 4.78 mmol) in acetonitrile (10 mL). The reaction was stirredat ambient temperature for 17.5 hours. The solvent was removed in vacuoand the resulting residue was dissolved in ethyl acetate and washed with1M NaOH (×4), water (×2), then brine (×1), and dried over Na₂SO₄.Filtration, followed by removal of the solvent in vacuo, gave a residuethat was purified via flash chromatography (1:4, ethyl acetate:hexanes,then 1:9 ethyl acetate:hexanes) to produce compound 104a (664 mg, 42%yield) as a colorless oil, ¹H NMR (CDCl₃, 300 MHz): 0.03 (s, 9H),0.96-1.02 (m, 2H), 1.80-1.95 (m, 4H), 2.29 (bs, 2H), 2.59-2.64 (m, 2H),3.46 (s, 2H), 4.15-4.29 (m, 4H), 7.21-7.30 (m, 5H); ESI (+) MS: 347(M+H⁺).

A mixture of compound 104a (645 mg, 1.86 mmol), Pd/C (250 mg, 10%, wet),and methanol (25 mL) was hydrogenated on a Parr apparatus at 50 psi H₂for 5 hours. The reaction was filtered through celite, and the solventremoved in vacuo to yield compound 104 (458 mg, 96% yield) as a whitesolid, ¹H NMR (CDCl₃, 300 MHz): 0.03 (s, 9H), 0.94-0.99 (m, 2H),1.76-1.91 (m, 5H), 2.60-2.64 (m, 2H), 2.94 (bs,2H), 4.11-4.18 (m, 4H);ESI (+) MS: 257 (M+H⁺), which was used with out further purification.

The Teoc precursor to compound 10 was prepared by the general couplingprocedure of Example 1 using compound 100 (805 mg, 0.880 mmol), amine104 (452 mg, 1.76 mmol), and manganese(IV) oxide (765 mg, 8.80 mmol),which provided a blue solid (875 mg, 94% yield), ESI (+) MS: 1055(M+H⁺). A mixture of this Teoc precursor (506 mg, 0.480 mmol),tetrabutylammonium fluoride (4.80 mL, 1.0 M in THF, 4.80 mmol), andacetonitrile (15 mL) was stirred in a sealed flask at 50° C. for 22hours. The reaction was cooled, and the solvent removed in vacuo. Theresidue was dissolved in chloroform and washed with water (×3) and driedover Na₂SO₄. Filtration followed by removal of the solvent in vacuoyielded a residue that was purified via MPLC (gradient, 2.5-10%methanol:methylene chloride), followed by reverse phase chromatography(C₁₈ silica, 9: 1, methanol:water), followed by preparatory thin layerchromatography (1.0 mm silica, 9: 1, methylene chloride:methanol, ˜50 mgper plate) to yield compound 10 as a blue solid (198 mg, 45% yield),Mp=210-212° C.; ESI (+) MS: 911 (M+H⁺); UV/Vis: λ_(max)=640.9 nm.

EXAMPLE 19

Synthesis of Compound No. 16 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 10 (87 mg, 0.0955 mmol) to provide compound16 (83 mg, 100% yield) of a blue solid, Mp=208-229° C.; ESI (+) MS: 869(M+H⁺); UV/Vis: λ_(max)=641.2 nm.

EXAMPLE 20

Synthesis of Compound No. 13 (see Table 1 for Structure)

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (6.68 g, 7.30 mmol),piperidin-4-yl-carbamic acid isobutyl ester (EP 467325, 2.92 g, 14.6mmol), and manganese(IV) oxide (6.35 g, 73.0 mmol) to provide compound13 (4.59 g, 63% yield) as a blue solid, Mp=184-188° C.; ESI (+) MS: 999(M+H⁺); UV/Vis: λ_(max)=643.1 nm.

EXAMPLE 21

Synthesis of Compound No. 14 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 13 (304 mg, 0.304 mmol) to provide compound14 (251 mg, 86%) as a blue solid, Mp=173-180° C.; ESI (+) MS: 957(M+H⁺); UV/Vis: λ_(max)=643.0 nm.

EXAMPLE 22

Synthesis of Compound No. 17 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 17 was preparedas follows:

1-(tert-Butoxycarbonyl)-4-piperidone (998 mg, 4.75 mmol) in methanol (15mL) was treated with dimethylamine hydrochloride (800 mg, 9.8 mmol) andsodium cyanoborohydride (270 mg, 4.3 mmol) at rt. After 4 days,concentrated HCl (˜10 mL) was added and volume of the reaction wasreduced in vacuo. The resulting residue was dissolved in H₂O (30 mL) andtreated with a 2M NaOH solution to achieve a pH of ˜10. The aqueoussolution was extracted with methylene chloride (3×20 mL) and thecombined organics were dried over Na₂SO₄ and removed in vacuo. Theresulting amine 106 (169 mg), ¹H NMR (CDCl₃, 300 MHz):δ3.14 (m, 2H),2.58 (td, J=12.3, 2.4 Hz, 2H), 2.28 (s, 6H), 2.22 (m, 1H), 1.82 (m, 2H),1.68 (s, 1H), 1.37 (tdd, J=12.2, 12.2, 4.1 Hz, 2H); ESI (+) MS: 129(M+H⁺), was used without further purification.

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (724 mg, 0.791 mmol), amine 106 (169 mg,1.32 mmol), and manganese(IV) oxide (857 mg, 9.9 mmol) to providecompound 17 (158 mg, 22%) as a blue solid, ESI (+) MS: 927 (M+H⁺);UV/Vis: λ_(max)=646.3 nm.

EXAMPLE 23

Synthesis of Compound No. 18 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 17 (89 mg, 0.096 mmol) to provide compound18 (70 mg, 82%) as a blue solid, Mp=214-216° C.; ESI (+) MS: 885 (M+H⁺);UV/Vis: λ_(max)=646.5 nm.

EXAMPLE 23

Synthesis of Compound No. 19

To a stirred solution of rifamycin (12.0 g, 17.3 mmol) in toluene wasadded N-methyl-1,2-phenylenediamine (2.10 g, 17.3 mmol) at roomtemperature. The resulting solution was heated at 65° C. for 48 h. Thesolvent was removed under vacuum. The residue was dissolved in ethanol(100 ml), and manganese(IV) oxide (14.8 g, 173 mmol) was added to thesolution at room temperature. The mixture was stirred for another 48 h,followed by filtration through celite, which was washed with methylenechloride (200 mL). The combined filtrates were washed with H₂O (2×200ml) and the volume reduced in vacuo. Combiflash column (1:19:180 MeOH,ethyl acetate, methylene chloride) afforded 107 (3.8 g, 27%) as a purplesolid, ESI (+) MS: 798 (M+H⁺).

EXAMPLE 24

Synthesis of Compound No. 19 (see Table 1 for Structure)

The title compound was prepared by the general coupling procedure ofExample 1 (heated to 50° C. for 5 days) using compound 107 (2.0 g, 2.5mmol), commercially available piperidin-4-yl-carbamic acid ethyl ester(2.15 g, 12.5 mmol), and manganese(IV) oxide (21.8 g, 25 mmol) toprovide compound 19 (570 mg, 23% yield) as a blue solid, Mp=220-240° C.;ESI (+) MS: 968 (M+H⁺); UV/Vis: λ_(max)=592.7 nm.

EXAMPLE 25

Synthesis of Compound No. 31 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 19 (510 mg, 0.526 mmol) to provide compound31 (120 mg, 24%) as a blue solid, Mp=250-252° C.; ESI (+) MS: 926(M+H⁺); UV/Vis: λ_(max)=591.5 nm.

EXAMPLE 26

Synthesis of Compound No. 21 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 21 was preparedas follows:

Ethyl chloroformate (0.171 mL, 1.79 mmol) was added to a stirredsolution of compound 101b (306 mg, 1.63 mmol), triethylamine (0.341 mL,2.45 mmol), and methylene chloride (15 mL) at 0° C. The reaction wasstirred under an atmosphere of N₂ at 0° C. for 30 minutes. The reactionwas quenched with sat. NaHCO₃ and the aqueous layer was extracted withmethylene chloride (×1). The organics were combined and dried overNa₂SO₄. Filtration followed by removal of the solvent in vacuo yielded aresidue that was purified via flash chromatography (19:1, methylenechloride:methanol) to yield compound 108a (381 mg, 90% yield) as a whitesolid,. ¹H NMR (CDCl₃, 300 MHz): 1.23 (t,j=7.0 Hz, 3H), 1.50 (s, 2H),2.38 (d,j=8.4 Hz, 2H), 2.93 (s, 1H), 3.06 (d,j=8.8 Hz, 2H), 3.55 (s,2H), 4.10 (q,j=7.0 Hz, 2H), 4.67 (bs, 1H), 7.19-7.31 (m, 5H); ESI (+)MS: 261 (M+H⁺).

A mixture of compound 108a (375 mg, 1.44 mmol), Pd/C (250 mg, 10%, wet),and methanol (25 mL) was hydrogenated on a Parr apparatus at 50 psi H₂for 13 hours. The reaction was filtered through celite, and the solventremoved in vacuo to yield compound 108 (265 mg) as a light yellow oil,¹H NMR (CDCl₃, 300 MHz): 1.24 (t,j=7.0 Hz, 3H), 1.60 (s, 2H), 1.82 (s,1H), 2.36 (d,j=1.7 Hz, 1H), 2.94 (d,j=11.4 Hz, 2H), 3.16 (d,j=11.5 Hz,2H), 4.11 (q,j=6.9 Hz, 2H), 4.73 (bs, 1H); ESI (+) MS: 171 (M+H⁺), whichwas used without further purification.

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (659 mg, 0.720 mmol), amine 108 (245 mg,1.44 mmol), and manganese(IV) oxide (626 mg, 7.20 mmol) to providecompound 21 (129 mg, 18% yield) as a blue solid, Mp=>350° C.; ESI (+)MS: 969 (M+H⁺); UV/Vis: λ_(max)=639.7 nm.

EXAMPLE 27

Synthesis of Compound No. 22 (see Table 1 for Structure)

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (6.19 g, 6.70 mmol),piperidin-4-yl-carbamic acid isopropyl ester (EP 467325, 2.52 g, 13.5mmol), and manganese(IV) oxide (5.86 g, 67.0 mmol) to provide compound22 (2.39 g, 36% yield) as a blue solid, Mp=>300° C.; ESI (+) MS: 985(M+H⁺); UV/Vis: λ_(max)=643.0 nm.

EXAMPLE 28

Synthesis of Compound No. 29 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 22 (0.850 g, 0.860 mmol) to provide compound29 (0.640 g, 78%) as a blue solid, Mp=222-225° C.; ESI (+) MS: 911(M+H⁺); UV/Vis: λ_(max)=643.0 nm.

EXAMPLE 29

Synthesis of Compound No. 23 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 23 was preparedas follows:

4-Amino-I-benzylpiperidine (685 mg, 3.61 mmol) in methylene chloride (10mL) was treated with trifluoromethanesulfonic anhydride (1.6 mL, 3.4mmol) and triethylamine (0.8 mL, 5.7 mmol) at 0° C. After warming to rtovernight the reaction was added to a saturated NaHCO₃ solution (100 mL)and extracted with methylene chloride (3×25 mL). The combined organicswere dried over Na₂SO₄ and concentrated in vacuo. The resulting residuewas purified via MPLC (silica gel, gradient 0-5%, methanol in methylenechloride) to yield compound 109a (844 mg, 2.6 mmol, 73%). ESI (+) MS:323 (M+H⁺).

Compound 109a (844 mg, 2.62 mmol) was dissolved in methanol (15 mL) andtreated with HCl in diethylether (2 M, 1.5 mL). The solvent was removedunder reduced pressure and the residue was redissolved in methanol (50mL). Palladium hydroxide on carbon (20%, 152 mg, 0.22 mmol) was addedand the reaction was treated with H₂ (55 psi). After 15 h the reactionwas filtered (celite, vacuum) and concentrated in vacuo. The resultingamine salt 109 (620 mg), ¹H NMR (DMSO-d₆, 300 MHz): δ 10.15 (s, 1H),9.47 (s, 1H), 9.29 (s, 1H), 3.68 (m, 1H), 3.25 (m, 2H), 3.02 (m, 2H),1.95 (dd, J=13.4, 2.8 Hz, 2H), 1.76 (m, 2H); ESI (+) MS: 233 (M+H⁺), wasused without further purification.

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (1.58 g, 1.73 mmol), amine 109 (580 mg,2.16 mmol), manganese(IV) oxide (1.37 g, 15.7 mmol) and triethylamine(0.90 mL, 6.5 mmol) to provide compound 23 (351 mg, 20% yield) as a bluesolid, Mp=238-240° C.; ESI (+) MS: 1031 (M+H⁺); UV/Vis: λ_(max)=638.7nm.

EXAMPLE 30

Synthesis of Compound No. 38 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 23 (103 mg, 0.099 mmol) to provide compound38 (22 mg, 22% yield) as a blue solid, Mp=245-246° C.; APCI (+) MS: 989(M+H⁺); UV/Vis: λ_(max)=639.7 nm.

EXAMPLE 31

Synthesis of Compound No. 24 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 24 was preparedas follows:

Butyryl chloride (1.30 mL, 12.4 mmol) was added to a stirred solution of4-amino-1-benzylpiperidine (700 mg, 3.68 mmol), triethylamine (0.5 mL,3.6 mmol), 4-dimethylaminopyridine (5 mg, 0.04 mmol), methylene chloride(10 mL) at 0°C. After addition, the reaction was allowed to come to rtand stirred for 4 days. After removal of the solvents in vacuo, theresulting residue was purified via MPLC (silica gel, gradient 2.5-7.5%,methanol in methylene chloride) to yield compound 110a (408 mg, 1.57mmol, 42%). ESI (+) MS: 261 (M+H⁺).

A mixture of compound 110a (400 mg, 1.54 mmol), palladium hydroxide oncarbon (20%, 122 mg, 0.17 mmol), and methanol (25 mL) was hydrogenatedon a Parr apparatus at 55 psi H₂ for 2 days. The reaction was filteredthrough celite, and the solvent removed in vacuo to yield compound 110(345 mg), ¹H NMR (CD₃OD, 300 MHz): δ3.93 (m, 1H), 3.41 (m, 2H), 3.09 (m,2H), 2.20 (t, J=12.6 Hz, 2H), 2.09 (dd, J=13.8, 2.0 Hz, 2H), 1.70 (m,2H), 1.64 (m, 2H), 0.94 (t, J=7.4 Hz, 3H); ESI (+) MS: 171 (M+H⁺), whichwas used without further purification.

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (997 mg, 1.09 mmol), amine 110 (345 mg,1.67 mmol), manganese(IV) oxide (1.10 g, 12.6 mmol), and triethylamine(2.0 mL, 14 mmol) to provide compound 24 (291 mg, 28% yield) as a bluesolid, Mp=222-226° C.; ESI (+) MS: 969 (M+H⁺); UV/Vis: λ_(max)=643.2 nm.

EXAMPLE 32

Synthesis of Compound No. 36 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 24 (83 mg, 0.086 mmol) to provide compound36 (46 mg, 58% yield) as a blue solid, Mp=228-230° C.; APCI (+) MS: 927(M+H⁺); UV/Vis: λ_(max)=643.9 nm.

EXAMPLE 33

Synthesis of Compound No. 25 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 25 was preparedas follows:

Methanesulfonyl chloride (0.5 mL, 6.4 mmol) was added to a stirredsolution of 4-amino-1-benzylpiperidine (670 mg, 3.53 mmol),triethylamine (0.9 mL, 6.5 mmol), and methylene chloride (10 mL) at 0°C. After addition, the reaction stirred under an atmosphere of N₂ at 0°C. for 30 minutes, warmed to rt, and stirred an additional 32 h. Afterremoval of the solvents in vacuo, the resulting residue was purified viaMPLC (silica gel, gradient 0-10%, methanol in methylene chloride) toyield compound 111a (844 mg, 3.15 mmol, 89% yield), ESI (+) MS: 269(M+H⁺).

Compound 111a (460 mg, 1.72 mmol) was dissolved in methylene chloride(10 mL) and treated with 1-chloroethyl chloroformate (0.23 mL, 2.1 mmol)at 0° C. After 15 minutes the reaction was heated to reflux for 2 h. Thesolvent was removed under reduced pressure and the residue was dissolvedin methanol (10 mL) and heated at reflux for 1 h. The reaction wasreduced in vacuo and the resulting mixture of compound 111a and aminesalt 111 was used without further purification. ESI (+) MS: 179(M+H⁺).

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (1.43 g, 1.56 mmol), amine 111 (413 mg,1.93 mmol), manganese(IV) oxide (1.01 g, 11.6 mmol), and triethylamine(2.0 mL, 14 mmol) to provide compound 25 (390 mg, 26% yield) as a bluesolid, Mp=256-257° C.; ESI (+) MS: 977 (M+H⁺); UV/Vis: λ_(max)=642.0 nm.

EXAMPLE 34

Synthesis of Compound No. 27 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 25 (98 mg, 0.10 mmol) to provide compound 27(78 mg, 83% yield) as a blue solid, Mp=270-273° C.; ESI (+) MS: 935(M+H⁺); UV/Vis: λ_(max)=641.0 nm.

EXAMPLE 35

Synthesis of Compound No. 28 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 28 was preparedas follows:

4-Amino-1-benzylpiperidine (1.01 g, 5.3 mmol) in methylene chloride (10mL) was treated with N-propyl isocyanate (2.5 mL, 26.7 mmol) anddiisopropylethylamine (3.5 mL, 20.1 mmol) at rt. After 16 h the reactionwas added to a saturated NaHCO₃ solution (100 mL) and extracted withmethylene chloride (3×25 mL). The combined organics were dried overNa₂SO₄ and reduced in vacuo. The resulting residue was purified via MPLC(silica gel, gradient 2.5-12.5%, methanol in methylene chloride) toyield compound 112a (1.13 g, 4.12 mmol, 78% yield), ESI (+) MS: 276(M+H⁺).

A mixture of compound 112a (1.13 g, 4.12 mmol), palladium hydroxide oncarbon (20%, 325 mg, 0.46 mmol), and methanol (25 mL) was hydrogenatedon a Parr apparatus at 55 psi H₂ for 4 days. The reaction was filteredthrough celite, and the solvent removed in vacuo to yield compound 112(862 mg), ¹H NMR (CD₃OD, 300 MHz): δ 3.72 (m, 1H), 3.34 (m, 2H),3.09-3.04 (m, 2H), 3.05-2.97 (m, 2H), 2.06 (dd, J=13.8, 2.8 Hz, 2H),1.59 (m, 2H), 1.48 (m, 2H), 0.91 (t, J=7.4 Hz, 3H), which was usedwithout any further purification.

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (1.68 g, 1.84 mmol), amine 112 (862 mg, 3.9mmol), manganese(IV) oxide (2.0 g, 23 mmol), and triethylamine (1.0 mL,7.2 mmol) to provide compound 28 (1.27 g, 70% yield) as a blue solid,Mp=260-263° C.; ESI (+) MS: 984 (M+H⁺); UV/Vis: λ_(max)=644.5 nm.

EXAMPLE 36

Synthesis of Compound No. 26 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 28 (300 mg, 0.305 mmol) to provide compound26 (187 mg, 65% yield) as a blue solid, Mp=265-266° C.; ESI (+) MS: 942(M+H⁺); UV/Vis: λ_(max)=644.0 nm.

EXAMPLE 37

Synthesis of Compound No. 32 (see Table 1 for Structure)

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (2.25 g, 2.76 mmol),piperidin-4-yl-carbamic acid methyl ester (873 mg, 5.52 mmol, Biorg.Med. Chem Lett. 2001, 2475-2480), and manganese(IV) oxide (2.40 g, 27.6mmol) to provide compound 32 (2.27 g, 86% yield) as a blue solid,Mp=149-153° C.; ESI (+) MS: 957 (M+H⁺); UV/Vis: λ_(max)=645.5 nm.

EXAMPLE 38

Synthesis of Compound No. 30 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 32 (438 mg, 0.458 mmol) to provide compound30 (386 mg, 92%) as a blue solid, Mp=212-225° C.; ESI (+) MS: 915(M+H⁺); UV/Vis: λ_(max)=643.1 nm.

EXAMPLE 39

Synthesis of Compound No. 33 (see Table 1 for Structure)

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (1.20 g, 1.31 mmol), 4-aminopiperidine(0.45 mL, 4.3 mmol), and manganese(IV) oxide (774 mg, 8.90 mmol) toprovide compound 33 (431 mg, 37% yield) as a blue solid, Mp=278-279° C.;ESI (+) MS: 899 (M+H⁺); UV/Vis: λ_(max)=647.7 nm.

EXAMPLE 40

Synthesis of Compound No. 39 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 33 (110 mg, 0.122 mmol) to provide 39 (95mg, 91% yield) as a blue solid, Mp=265-266° C.; ESI (+) MS: 857 (M+H⁺);UV/Vis: λ_(max)=647.8 nm.

EXAMPLE 41

Synthesis of Compound No. 34 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 34 was preparedas follows:

4-Amino-1-benzylpiperidine (990 mg, 5.21 mmol) in methylene chloride (10mL) was treated with N-ethyl isocyanate (2.0 mL, 25 mmol), anddiisopropylethylamine (1.8 mL, 10 mmol) at rt. After 16 h the reactionwas added to a saturated NaHCO₃ solution (100 mL) and extracted withmethylene chloride (3×25 mL). The combined organics were dried overNa₂SO₄ and reduced in vacuo. The resulting residue was purified via MPLC(silica gel, gradient 2.5-12.5%, methanol in methylene chloride) toyield compound 113a (1.14 g, 4.39 mmol, 84% yield), ESI (+) MS: 262(M+H⁺).

A mixture of compound 113a (1.06 g, 4.08 mmol), palladium hydroxide oncarbon (20%, 350 mg, 0.50 mmol), and methanol (50 mL) was hydrogenatedon a Parr apparatus at 55 psi H₂ for 4 hours. The reaction was filteredthrough celite, and the solvent removed in vacuo to yield a mixture ofcompounds 113a and 113, ¹H NMR (CD₃OD, 300 MHz): 6 3.79 (m, 1H), 3.41(m, 2H), 3.22-3.15 (m, 2H), 3.15-3.05 (m, 2H), 2.09 (m, 2H), 1.73 (m,2H), 1.13 (t, J=7.2 Hz, 3H), which were used without any furtherpurification.

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (2.00 g, 2.16 mmol), amine 113 (1.05 g,5.07 mmol), manganese(IV) oxide (2.20 g, 25 mmol) and triethylamine (1.5mL, 11 mmol) to provide compound 34 (350 mg (17%) as a blue solid,Mp=234-236° C.; ESI (+) MS: 970 (M+H⁺); UV/Vis: λ_(max)=644.6 nm.

EXAMPLE 42

Synthesis of Compound No. 37 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 34 (200 mg, 0.206 mmol) to provide compound37 (116 mg, 61%) as a blue solid, Mp=321-322° C.; APCI (+) MS: 928(M+H⁺); UV/Vis: λ_(max)=645.0 nm.

EXAMPLE 43

Synthesis of Compound No. 35 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 35 was preparedas follows:

Propanesulfonyl chloride (0.9 mL, 7.9 mmol) was added to a stirredsolution of 4-amino-1-benzylpiperidine (890 mg, 4.68 mmol),triethylamine (0.8 mL, 5.7 mmol), and methylene chloride (10 mL) at 0°C. After addition, the reaction stirred under an atmosphere of N₂ at 0°C. for 30 minutes, warmed to rt, and stirred an additional 32 h. Afterremoval of the solvents in vacuo, the resulting residue was purified viaMPLC (silica gel, gradient 0-5%, methanol in methylene chloride) toyield compound 114a (1.02 g, 3.44 mmol, 89%), ¹H NMR (CDCl₃, 300MHz):δ7.34-7.25 (m, 5H), 4.05 (d, J=7.7 Hz, 1H), 3.49 (s, 2H), 3.31 (m,1H), 2.98 (m, 2H), 2.80 (m, 2H), 2.11 (m, 2H), 1.99 (m, 2H), 1.84 (m,2H), 1.58 (m, 2H), 1.05 (t, J=7.4 Hz, 3H); ESI (+) MS: 297 (M+H⁺).

Compound 114a (883 mg, 2.98 mmol) was dissolved in methylene chloride(10 mL) and treated with 1-chloroethyl chloroformate (0.23 mL, 2.1 mmol)at 0° C. After 15 minutes the reaction was heated to reflux for 2 h. Thesolvent was removed under reduced pressure and the residue was dissolvedin methanol (10 mL) and heated at reflux for 1h. The reaction wasreduced in vacuo and the resulting compound 114 (769 mg) was usedwithout further purification.

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (1.24 g, 1.36 mmol), amine 114 (769 mg,3.17 mmol), manganese(IV) oxide (1.40 g, 16 mmol), and triethylamine(0.8 mL, 6 mmol) to provide compound 35 (640 mg, 47% yield) as a bluesolid, Mp=220-226° C.; ESI (+) MS: 1005 (M+H⁺); UV/Vis: λ_(max)=641.7nm.

EXAMPLE 44

Synthesis of Compound No. 67 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 35 (198 mg, 0.197 mmol) to provide compound67 (110 mg, 58% yield) as a blue solid, Mp=215-216° C.; APCI (+) MS: 963(M+H⁺); UV/Vis: λ_(max)=641.5 nm.

EXAMPLE 45

Synthesis of Compound No. 40 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 40 was preparedas follows:

Ethyl chloroformate (0.491 mL, 5.14 mmol) was added to a stirredsolution of compound 102b (1.01 g, 4.67 mmol, U.S. Pat. No. 5,654,318),triethylamine (0.980 mL, 7.01 mmol), and methylene chloride (20 mL) at0° C. The reaction was stirred under an atmosphere of N₂ at 0° C. for 30minutes, quenched with sat. NaHCO₃ and the aqueous layer extracted withmethylene chloride (×1). The organics were combined and dried overNa₂SO₄. Filtration, followed by removal of the solvent in vacuo, yieldeda residue that was purified via flash chromatography (1:1, ethyl acetatehexanes) to yield compound 115a (1.06 g, 79% yield) as a colorless oil,¹H NMR (CDCl₃, 300 MHz): 1.24 (t,j=7.1 Hz, 3H), 1.37-1.72 (m, 4H),2.10-2.16 (m, 1H), 2.51 (d,j=8.9 Hz, 1H), 2.64 (t,j=8.2 Hz, 1H),2.75-2.84 (m, 3H), 3.66 (dd,j=24.2, 13.1 Hz, 2H), 3.92 (bs, 1H), 4.11(q,j=7.1 Hz, 2H), 4.63 (bs, 1H), 7.22-7.32 (m, 5H); ESI (+) MS: 289(M+H⁺).

A mixture of compound 115a (1.05 g, 3.64 mmol), Pd/C (300 mg, 10%, wet),and methanol (25 mL) was hydrogenated on a Parr apparatus at 50 psi H₂for 14.5 hours. The reaction was filtered through celite, and thesolvent removed in vacuo to yield compound 115 (767 mg, quant., wet withmethanol) as a light yellow oil, ¹H NMR (CDCl₃, 500 MHz): 1.27 (t,j=3.5Hz, 3H), 1.38-1.45 (m, 2H), 1.68-1.73 (m, 2H), 2.04-2.07 (m, 2H),2.75-2.83 (m, 3H), 3.09-3.18 (m, 2H), 4.00 (d,j=11.5 Hz, 1H), 4.11-4.17(m, 2H), 4.52-4.54 (m, 1H); ESI (+) MS: 199 (M+H⁺), which was used without further purification.

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (1.65 g, 1.80 mmol), amine 115 (712 mg,3.59 mmol) and manganese(IV) oxide (1.56 g, 18.0 mmol) to providecompound 40, Diastereomer A (555 mg (31%) as a blue solid (higher R_(ƒ)using ethyl acetate as eluent) and compound 40b, Diastereomer B (660 mg,37% yield, not shown in Table) as a blue solid (lower R_(ƒ)using ethylacetate as eluent), Mp=>300° C.; ESI (+) MS: 997 (M+H⁺); UV/Vis:λ_(max)=644.0 nm.

EXAMPLE 46

Synthesis of Compound No. 42 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 40 (110 mg, 0.110 mmol) to provide compound42 (97 mg, 92% yield) as a blue solid, as mixture of diastereomers (asdetermined by HPLC and ¹H NMR), Mp=>300° C.; ESI (+) MS: 955 (M+H⁺);UV/Vis: λ_(max)=644.0 nm.

EXAMPLE 47

Synthesis of Compound No. 41 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 41 was preparedas follows:

A stirred solution of 4-amino-1-benzylpiperidine (3.0 mL, 15.0 mmol) andtriethylamine (3.4 mL, 24.4 mmol) in methylene chloride (50 mL) wascooled to 0° C. using an ice bath. To the reaction flask was addedchloroformic acid, 2-methoxyethyl ester (2.00 mL, 17.5 mmol). Thereaction was stirred under nitrogen at 0° C. for 45 minutes, quenchedwith saturated sodium bicarbonate (150 mL), extracted with methylenechloride (×3), and concentrated in vacuo to provide an oily residue. Theresidue was purified by flash chromatography (1:19, methanol:methylenechloride) to yield 4.64 g of compound 116a as a pale yellow oil, ¹H NMR(300 MHz, CDCl₃) δ1.36-1.52 (m, 2H), 1.90 (d, 2H), 2.03-2.17 (t, 2H),2.78 (d, 2H), 3.38 (s, 3.44-3.64 (m, 5H), 4.13-4.31 (m, 2H), 4.69 (d,1H), 7.19-7.35 (m, 5H); ESI (+) MS: 293.

Compound 116a (4.64 g, 15.9 mmol) was dissolved in methanol (75 mL) andreacted with Pd/C (1.00 g, 10 wt %, wet) on a Parr apparatus at roomtemperature under 50 psi of hydrogen for 16 hours. The reaction wasfiltered through celite and the solids rinsed with methanol (250 mL).The filtrate and rinse were concentrated down to provide 3.13 g (97%yield) of amine 116 as a light yellow oil. ¹H NMR (300 MHz, CDCl₃) δ1.23-1.40 (m, 2H), 1.93 (d, 2H), 2.08 (s, 2H), 2.57-2.74 (t, 2H), 3.06(d, 2H), 3.38 (s, 3H), 3.49-3.67 (m, 2H), 4.12-4.32 (m, 2H), 4.85 (s,1H). ESI (+) MS: 203.

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (4.50 g, 5.16 mmol), amine 116 (3.13 g,15.5 mmol), and manganese(IV) oxide (4.60 g, 52.9 mmol) to providecompound 41 (4.34 g, 84%) as a blue solid, Mp=175-183° C.; ESI (+) MS:1001 (M+H⁺); UV/Vis: λ_(max)=642.9 nm.

EXAMPLE 48

Synthesis of Compound No. 44 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 44 was preparedas follows:

4-Amino-1-benzylpiperidine (1.31 g, 6.89 mmol) in methylene chloride (20mL) was treated with acetic anhydride (0.80 mL, 8.5 mmol) andtriethylamine (2.0 mL, 14 mmol) at rt. After 24 h the reaction was addedto a saturated NaHCO₃ solution (100 mL) and extracted with methylenechloride (3×25 mL). The combined organics were dried over Na₂SO₄ andreduced in vacuo. The resulting residue was purified via MPLC (silicagel, gradient 2.5-12.5%, methanol in methylene chloride) to yieldcompound 117a (1.35 g, 5.8 mmol, 84% yield), ESI (+) MS: 233 (M+H⁺).

A mixture of compound 117a (1.35 g, 5.8 mmol), palladium on carbon (10%,421 mg, 0.79 mmol), and methanol (25 mL) was hydrogenated on a Parrapparatus at 55 psi H₂ for 24 hours. The reaction was filtered throughcelite, and the solvent removed in vacuo to yield compound 117, ¹H NMR(CDCl₃, 300 MHz): δ5.45 (s, 1H), 3.87 (m, 1H), 3.07 (m, 2H), 2.69 (td,J=12.2, 2.5 Hz, 2H), 1.97 (s, 3H), 195 (m, 2H), 1.75 (s, 1H), 1.31 (m,2H), which was used without any further purification.

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (1.83 g, 2.0 mmol), compound 17 (950 mg,6.7 mmol), and manganese(IV) oxide (1.09 g, 12.5 mmol) to providecompound 44 (926 mg, 49% yield) as a blue solid, Mp=232-234° C.; ESI (+)MS: 941 (M+H⁺); UV/Vis: λ_(max)=643.6 nm.

EXAMPLE 49

Synthesis of Compound No. 43 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 44 (272 mg, 0.289 mmol) to provide compound43 (208 mg, 80%) as a blue solid, Mp=232-236° C.; ESI (+) MS: 899(M+H⁺); UV/Vis: λ_(max)=643.9 nm.

EXAMPLE 50

Synthesis of Compound No. 47 (see Table 1 for Structure)

The precursor amine used in the preparation of compound zz was preparedas follows:

Acetyl chloride (0.373 mL, 5.24 mmol) was added to a stirred solution ofcompound 102b (1.03 g, 4.76 mmol), triethylamine (1.0 mL, 7.14 mmol),and methylene chloride (20 mL) at 0° C. The reaction was stirred underan atmosphere of N₂ at 0° C. for 1 hour. The reaction was quenched withsat. NaHCO₃, and the aqueous layer was extracted with methylene chloride(×1). The organics were combined and dried over Na₂SO₄. Filtrationfollowed by removal of the solvent in vacuo yielded a residue that waspurified via flash chromatography (9:1, methylene chloride:methanol) toyield compound 118a (1.09 g, 87%) as a light yellow oil, ¹H NMR (CDCl₃,300 MHz): 1.31-1.78 (m, 4H), 2.06-2.39 (m, 4H), 2.48-2.90 (m, 4.5H,conformer), 3.06-3.16 (m, 0.5H, conformer), 3.58-3.75 (m, 2.5H,conformer), 4.32-4.44 (m, 1H), 5.03 (q,j=8.5 Hz, 0.5H, conformer),7.21-7.35 (m, 5H); ESI (+) MS: 259 (M+H⁺).

A mixture of compound 118a (2.18 g, 8.43 mmol), Pd/C (500 mg, 10%, wet),and methanol (50 mL) was hydrogenated on a Parr apparatus at 50 psi H₂for 15 hours. The reaction was filtered through celite, and the solventremoved in vacuo to yield compound 118 (1.55 g, quant., wet withmethanol) as a light yellow oil, ¹H NMR (CDCl₃, 500 MHz): 1.35-1.53 (m,2H), 1.73-1.77 (m, 2H), 2.08-2.14 (m, 4H), 2.56 (t,j=10.0 Hz, 0.4H,conformer), 2.77 (t,j=10.4 Hz, 0.6H, conformer), 2.82-2.86 (m, 2H), 2.96(t,j=10.0 Hz, 0.4H, conformer), 2.82-2.86 (m, 2H), 3.05-3.09(m, 1H),3.16-3.25 (m, 1.6H, conformer), 3.66 (d,j=12.7 Hz, 0.6H, conformer),4.27 (q, j=8.1 Hz, 0.4H, conformer), 4.51 (d,j=13.9 Hz, 0.4H,conformer), 4.97 (q,j=8.1 Hz, 0.6H, conformer); ESI (+) MS: 169 (M+H⁺),that was used with out further purification.

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (3.81 g, 4.16 mmol), amine 118 (1.40 g,8.32 mmol) and manganese(IV) oxide (3.62 g, 41.6 mmol) to providecompound 47 (916 mg, 23% yield) as a blue solid, Mp=148-149° C.; ESI (+)MS: 967 (M+H⁺); UV/Vis: λ_(max)=644.0 nm.

EXAMPLE 51

Synthesis of Compound No. 46 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 47 (214 mg, 0.221 mmol) to provide compound46 (178 mg, 87% yield) as a blue solid, Mp=148-149° C.; ESI (+) MS: 967(M+H⁺); UV/Vis: λ_(max)=644.0 nm.

EXAMPLE 52

Synthesis of Compound Nos. 45 and 49 (see Table 1 for Structure)

The precursor amine used in the preparation of compounds 45 and 49 wasprepared as follows:

Methylthiolchloroformate (1.0 g, 9.04 mmol) was added to a stirredsolution of commercially available 4-amino-1-benzylpiperidine (1.68 ML,8.22 mmol), triethylamine (1.71 mL, 12.3 mmol), and methylene chloride(30 mL) at 0° C. The reaction was stirred at 0° C. under an atmosphereof N₂ for 30 minutes, quenched with sat. NaHCO₃ and the aqueous layerwas extracted once with methylene chloride. The organics were combinedand dried over Na₂SO₄. Filtration, followed by removal of the solvent invacuo, yielded a residue that was purified via flash chromatography(3:2, ethyl acetate:hexanes) to yield compound 119a (2.17 g, 100%) as awhite solid, ¹H NMR (CDCl₃, 300 MHz): 1.41-1.53 (m, 2H), 1.93 (d,j=11.6Hz, 2H), 2.06-2.15 (m, 2H), 2.34 (s, 3H), 2.79 (d,j=11.9 Hz, 2H), 3.79(s, 2H), 3.79 (bs, 1H), 5.22 (bs, 1H), 7.23-7.34 (m, 5H); ESI (+) MS:265 (M+H⁺).

α-Chloroethylchloroformate (0.870 mL, 8.06 mmol) was added slowly (overa period of 15 minutes) to a stirred solution of 119a (2.13 g, 8.06mmol) in methylene chloride (20 mL) cooled in a brine/ice bath. Thebrine/ice bath was removed, and the reaction was refluxed with stirringfor 1.5 hours. The reaction was cooled to ambient temperature andmethanol (6 mL) was added. The reaction was again refluxed with stirringfor 2 hours. The reaction was cooled to ambient temperature and thesolvent was removed in vacuo. The resulting residue was purified viaflash chromatography (1:9:40, NH₄OH:methanol:methylene chloride) toyield compound 119 (1.07 g, 74%) as a yellow semi-solid, ¹H NMR (CDCl₃,300 MHz): 1.34-1.46 (m, 2H), 1.96-2.00 (m, 2H), 2.34 (s, 3H), 2.66-2.75(m, 2H), 3.08-3.14 (m, 2H), 3.88-3.92 (m, 1H), 5.62 (bs, 1H); ESI (+)MS: 175 (M+H⁺), which was used without further purification.

The title compounds were prepared by the general coupling procedure ofExample 1 using compound 100 (2.68 g, 2.93 mmol), amine 119 (1.02 g,5.85 mmol), and manganese(IV) oxide (2.55 g, 29.3 mmol) to provide 859mg of a blue solid (impure compound 45, higher R_(ƒ) using ethyl acetateas an eluent) and 603 mg of a blue solid (impure compound 49, lowerR_(ƒ) using ethyl acetate as an eluent) after flash chromatography.

Impure compound 45 was purified via MPLC (gradient, 2-20%tetrahydrofuran:methylene chloride, followed by gradient, 1.25-2.5%methanol:methylene chloride) to provide compound 45 (162 mg, 6% yield),Mp=150-155° C.; ESI (+) MS: 973 (M+H⁺); UV/Vis: λ_(max)=642.0 nm.

Impure compound 49 was purified via MPLC (gradient, 1.25-4.5%methanol:methylene chloride) followed by preparatory thin layerchromatography (1.0 mm silica, ethyl acetate, ˜50 mg per plate, eachplate resolved 6 times) to provide compound 49 (87 mg), Mp=>300° C.; ESI(+) MS: 1099 (M+H⁺); UV/Vis: λ_(max)=643.1 nm.

EXAMPLE 53

Synthesis of Compound No. 48 (see Table 1 for Structure)

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (5.20 g, 5.96 mmol),4-(tert-butoxycarbonylamino)piperidine (240 mg, 11.9 mmol) andmanganese(IV) oxide (5.20 g, 59.8 mmol) to provide 2.15 g (36%) of ablue solid, Mp=218-228° C.; ESI (+) MS: 999 (M+H⁺); UV/Vis:λ_(max)=643.1 nm.

EXAMPLE 54

Synthesis of Compound No. 52 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 48 (0.300 g, 0.300 mmol) to provide compound52 (0.273 g, 95%) as of a blue solid, Mp=230-232° C.; ESI (−) MS: 956(M+H⁺); UV/Vis: λ_(max)=643.5 nm.

EXAMPLE 55

Synthesis of Compound No. 50 (see Table 1 for Structure)

A mixture of compound 45 (397 mg, 0.480 mmol), N-methylpiperazine (0.230mL, 2.04 mmol) and methyl sulfoxide (5 mL) was stirred at ambienttemperature for 6 days. The reaction was diluted with methylene chlorideand poured over water. The aqueous layer was extracted with methylenechloride (×2) and the combined organics were washed with water (×2) thenbrine (×1) and dried over Na₂SO₄. Filtration followed by removal of thesolvent in vacuo yielded a residue that was purified via MPLC (gradient,2.5-10% methanol:methylene chloride) to yield compound 50 as a bluesolid (74 mg, 18% yield). Mp=>300° C., ESI (+) MS: 1025 (M+H⁺), UV/Vis:λ_(max)=644.9 nm. Example 56

Synthesis of Compound No. 51 (see Table 1 for Structure)

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (1.00 g, 1.10 mmol),(piperidin-4-ylmethyl)-carbamic acid ethyl ester (506 mg, 2.71 mmol),and manganese(IV) oxide (1.20 g, 13.80 mmol) to provide compound 51 (35mg, 3% yield) as a blue solid, Mp=175-185° C.; ESI (+) MS: 985 (M+H⁺);UV/Vis: λ_(max)=647.5 nm.

EXAMPLE 57

Synthesis of Compound No. 60 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 51 (117 mg, 0.11 mol) to provide compound 60(54 mg, 48%) as a blue solid, Mp=190-195° C.; ESI (+) MS: 943 (M+H⁺);UV/Vis: λ_(max)=638.9 nm.

EXAMPLE 58

Synthesis of Compound No. 53 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 53 was preparedas follows:

Formaldehyde (21.5 mL, 37 wt. % solution in water, 276 mmol) followed bysodium cyanoborohydride (2.89 g, 46.0 mmol) was added to a stirredsolution of compound 101b (1.73 g, 9.19 mmol) in acetonitrile (175 mL).The reaction was stirred under an atmosphere of N₂ at ambienttemperature for 15 minutes at which time the pH was adjusted to neutral(pH paper) with acetic acid. The reaction was stirred under anatmosphere of N₂ at ambient temperature for 75 minutes (adjusting pHevery 15 minutes if necessary), and the solvent was removed in vacuo.Aqueous NaOH (175 mL, 2M) was added, and the aqueous layer extractedwith diethyl ether (×5). The organics were combined and dried overNa₂SO₄. Filtration followed by removal of the solvent in vacuo yielded aresidue that was purified via flash chromatography (19:1 to 9:1,methylene chloride:methanol) to yield compound 120a (1.08 g, 57% yield)as a yellow oil, ¹H NMR (CDCl₃, 300 MHz): 1.42-1.44 (m, 2H), 1.95-1.97(m, 1H), 2.30 (s, 6H), 2.39 (d,j=8.5 Hz, 2H), 2.94 (d,j=8.8 Hz, 2H),3.56 (s, 2H), 7.22-7.32 (m, 5H); ESI (+) MS: 217 (M+H⁺).

A mixture of compound 120a (1.07 g, 4.95 mmol), Pd/C (300 mg, 10%, wet),and methanol (50 mL) was hydrogenated on a Parr apparatus at 50 psi H₂15hours. The reaction was filtered through celite, and the solvent removedin vacuo. The resulting residue was dissolved in methanol (50 mL) towhich was added Pd/C (600 mg, 10%, wet). The reaction was hydrogenatedon a Parr apparatus at 55 psi H₂, at 50° C. for 20.5 hours. The reactionwas filtered through celite, and the solvent removed in vacuo to yieldcompound 120 (528 mg, 84%) as a white solid. ¹H NMR (CDCl₃, 500 MHz):1.37 (t,j=2.0 Hz, 1H), 1.51 (q,j=2.0 Hz, 2H), 1.97 (bs, 1H), 2.29 (s,6H), 2.90 (d,j=11.5 Hz, 2H), 3.00 (d,j=11.0 Hz, 2H). ESI (+) MS: 127(M+H⁺).

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (1.88 g, 2.05 mmol), amine 120 (518 mg,4.10 mmol), and manganese(IV) oxide (1.78 g, 20.5 mmol) to providecompound 53 (1.24 g, 65% yield) as a blue solid, Mp=>370° C.; ESI (+)MS: 925 (M+H⁺); UV/Vis: λ_(max)=642.4 nm.

EXAMPLE 59

Synthesis of Compound No. 54 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 53 (347 mg, 0.375 mmol) to provide compound54 (289 mg, 87%) as a blue solid, Mp=>330° C.; ESI (+) MS: 883 (M+H⁺);UV/Vis: λ_(max)=642.2 nm.

EXAMPLE 60

Synthesis of Compound No. 55 (see Table 1 for Structure)

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (1.00 g, 1.10 mmol),N-(piperidin-4-ylmethyl)-acetamide (452 mg, 2.89 mmol, U.S. Pat. No.4,370,328), and manganese(IV) oxide (1.2 g, 13.80 mol) to providecompound 55 (215 mg, 20% yield) as a blue solid, Mp=214-218° C.; ESI (+)MS: 955 (M+H⁺); UV/Vis: λ_(max)=647.5 nm.

EXAMPLE 61

Synthesis of Compound No. 56 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 55 (152 mg, 0.15 mol) to provide compound 56(68 mg, 46% yield) as a blue solid, Mp=205-213° C.; ESI (+) MS: 913(M+H⁺); UV/Vis: λ_(max)=647.2 nm.

EXAMPLE 62

Synthesis of Compound No. 57 (see Table 1 for Structure)

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (2.70 g, 3.10 mmol), 4-phenylpiperidine(1.00 g, 6.20 mmol) and manganese(IV) oxide (2.70 g, 31.0 mmol) toprovide compound 57 (1.97 g, 66% yield) as a blue solid, Mp=220-222° C.;ESI (+) MS: 960 (M+H⁺); UV/Vis: λ_(max)=646.5 nm.

EXAMPLE 63

Synthesis of Compound No. 58 (see Table 1 for Structure)

The precursor amine used in the preparation of compound 58 was preparedas follows:

Formaldehyde (11.8 mL, 37 wt. % solution in water, 152 mmol) followed bysodium cyanoborohydride (1.59 g, 25.3 mmol) was added to a stirredsolution of compound 102b (2.19 g, 10.1 mmol) in acetonitrile (100 mL).The reaction was stirred under an atmosphere of N₂ at ambienttemperature for 15 minutes at which time the pH was adjusted to neutral(pH paper) with acetic acid. The reaction was stirred under anatmosphere of N₂ at ambient temperature for 4.5 hours, and the solventwas removed in vacuo. Aqueous NaOH (100 mL, 2M) was added, and extractedwith diethyl ether (×5). The organics were combined and dried overNa₂SO₄. Filtration followed by removal of the solvent in vacuo yielded aresidue that was purified via flash chromatography (9:1, methylenechloride:methanol) to yield compound 121a (1.95 g, 84% yield) as ayellow oil, ¹H NMR (CDCl₃, 300 MHz): 1.47-1.84 (m, 4H), 2.04-2.13 (m,1H), 2.19 (s, 3H), 2.23-2.33 (m, 1H), 2.57-2.66 (m, 3H), 2.72-2.81 (m,2H), 2.95-3.01 (m, 1H), 3.75 (s, 2H), 7.22-7.38, (m, 5H); ESI (+) MS:231 (M+H⁺).

A mixture of compound 121a (1.94 g, 8.42 mmol), Pd/C (500 mg, 10%, wet),and methanol (50 mL, sparged with H₂) was stirred under balloon pressureH₂ for 12 hours. The reaction was filtered through celite, and thesolvent removed in vacuo. The resulting residue was dissolved inmethanol (50 mL) to which was added Pd/C (500 mg, 10%, wet). Thereaction was hydrogenated on a Parr apparatus at 55 psi H₂, at 50° C.for 15.5 hours. The reaction was filtered through celite, and thesolvent removed in vacuo. The resulting residue was dissolved inmethanol (50 mL) to which was added Pd(OH)₂/C (500 mg, 20%, wet) and HCl(2 mL, conc.). The reaction was hydrogenated on a Parr apparatus at 50psi H₂ for 10 hours. The reaction was filtered through celite, and thesolvent removed in vacuo to yield compound 121 (1.70 g, 95% yield) as anoff-white foam, ¹H NMR (CD₃OD, 500 MHz): 1.86-2.01 (m, 4H), 2.86-2.92(m, 5H), 3.04-3.12 (m, 1H), 3.51-3.65 (m, 3H), 3.78 (d,j=10.5 Hz, 1H),3.91-3.94 (m, 2H). ESI (+) MS: 141 (M−2HCl+H⁺).

The title compound was prepared by the general coupling procedure ofExample 1 using compound 100 (6.68 g, 7.30 mmol), amine 121 (1.68 g,7.88 mmol), triethylamine (3.30 mL, 23.6 mmol), and manganese(IV) oxide(3.43 g, 39.4 mmol) to provide compound 58 (1.57 g, 42% yield) as a bluesolid after chromatography, Mp=>300° C.; ESI (+) MS: 939 (M+H⁺); UV/Vis:λ_(max)=649.5 nm. The other diastereomers was also obtained.

EXAMPLE 64

Synthesis of Compound No. 59 (see Table 1 for Structure)

The title compound was prepared by the general deacetylation procedureof Example 2 using compound 58 (335 mg, 0.357 mmol) to provide compound59 (278 mg, 87%) as a blue solid, Mp=>300° C.; ESI (+) MS: 897 (M+H⁺);UV/Vis: λ_(max)=649.4 nm.

EXAMPLE 65

Synthesis of25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-21,23-(1-methylethylideneacetal) rifamycin (compound 122)

The title compound was prepared as follows:

Following the procedure of Kump, et al., Helv. Chim. Acta. 56:2323(1973), 5 microliters of concentrated sulfuric acid is added to asolution of rifamycin S (1.1 g), dimethoxypropane (1.1 mL), and dryacetone (12 mL). The reaction mixture is stirred 45 minutes at roomtemperature. Anhydrous sodium carbonate (1 g) is added and stirringcontinued for 5 min. The solution is filtered and evaporated to dryness.The residue is purified by flash-chromatography; elution with 1%methanol in dichloromethane affords rifamycin S,cyclic-21,23-(1-methylethylidene acetal), compound, which is dissolvedin a cold solution of 5% NaOH in methanol (100 mL). The resultingmixture is stirred 18 hours at room temperature then diluted with icywater (100 mL), acidified (about pH 4) with citric acid and extractedwith dichloromethane (3×100 mL). The combined extracts are dried andevaporated to dryness. The residue, by crystallization from ethylether/petroleum ether, gives compound 122, 25-O-deacetyl-rifamycin S,cyclic-21,23-(1-methylethylidene acetal).

In addition to its use in the synthesis of25-(2″,3″-dihydroxypropylcarbonoxy)oxybenzoxazinorifamycin analogs, asdescribed below, compound 122 can also be used as a protectedintermediate for the synthesis of other analogs in which the 25 positionof rifamycin is derivatized. The conversion hydroxy groups to othermoieties is well known to those skilled in the art.

EXAMPLE 66

Synthesis of 25-O-deacetyl-21,23-(1-methylethylideneacetal)-5′-[4-isobutyl-1-piperazinyl]benzoxazinorifamycin (compound 123)

The title compound was prepared as follows:

Compound 122 (0.30g) and 2-hydroxyaniline (0.060 g) in 10 mL of tolueneare stirred at room temperature for 12 days. After insoluble substancesare filtered off, the solvent is removed under reduced pressure, and theresidue dissolved in 7 mL of ethanol. To the solution is added 0.15 g ofmanganese dioxide and the mixture stirred at room temperature for 7hours. Manganese dioxide is filtered off by using a filter aid and thesolvent removed under reduced pressure. The residue is purified bysilica-gel column-chromatography [eluent: chloroform-methanol (99:1)] togive the title compound (compound 123)

EXAMPLE 67

Synthesis of 25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-21,23-(1-methylethylideneacetal)-5′-[4-isobutyl-1-piperazinyl]benzoxazinorifamycin (compound 124)

The title compound was prepared as follows:

Compound 123 (0.20 g) and1-O-chloroformyl-2,3-O-isopropylidene-D,L-glycerol (0.90 g, seeSeligson, et al., Anticancer Drugs 12: 305-13,2001) are dissolved in dryCH₂Cl₂ (4 mL) and cooled to −70° C. under argon. Pyridine (0.28 mL) isadded, the cold bath removed, and the reaction stirred at roomtemperature for 4.5 hours. The organics are washed with water and driedover magnesium sulfate. After filtration and concentration under reducedpressure, the resulting solution is purified by silica gelchromatography (CH₂Cl₂/acetone) to yield compound 124.

EXAMPLE 68

Synthesis of25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-5′-[4-isobutyl-1-piperazinyl]benzoxazinorifamycin,(compound 126)

The title compound was prepared as follows:

The general coupling procedure of Example 1 is used to prepare the titlecompound. To compound 124 (0.20 g) in 3 mL of dimethyl sulfoxide isadded 100 mg of 4-isobutylpiperazine and 0.2 g of manganese dioxide. Themixture stirred at room temperature for 5 days. The reaction mixture isdiluted by addition of ethyl acetate and insoluble substances arefiltered off. The filtrate is washed successively with water and with asaturated aqueous solution of sodium chloride, and dried over anhydroussodium sulfate. After the drying agent is filtered off, the solvent isremoved under reduced pressure. The residue is purified twice bysilica-gel column-chromatography [eluent: ethyl acetate/hexane, 1:1 to9:1 gradient] to give 25-O-deacetyl-25-(2 ″,3″-dihydroxypropylcarbonoxy)-21,23-(1-methylethylideneacetal)-5′-[4-isobutyl-1-piperazinyl]benzoxazinorifamycin (compound125).

Compound 125 is dissolved in THF (2 mL) and 3% (v/v) sulfuric acid/water(0.7 mL) is added. The reaction mixture is stirred for 16 hours at 40°C. After cooling, the reaction mixture is diluted with water (5 mL) andextracted with ethyl acetate (2×10 mL). The combined organics are driedover magnesium sulfate, filtered, and the volatiles removed underreduced pressure to give25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-5′-[4-isobutyl-1-piperazinyl]benzoxazinorifamycin,compound 126.

EXAMPLE 69

MIC Assay

MICs of candidate compounds of the invention can be determined, forexample, by the method of Lee et al., Am. Rev. Respir. Dis. 136:349(1987). To a BACTEC 12B vial (4 mL of 7H12B medium), 0.1 mL of a 10-folddilution of subculture of the test organisms in 7H9 medium (opticaldensity at 540 nm, 0.1) is inoculated and cultured at 37° C. until agrowth index (GI) of 999 is reached. Then the broth culture is removedand diluted 100-fold, and 0.1 mL of the dilution is inoculated into aBACTEC 12B vial with or without a candidate compound. The candidatecompound containing vials can hold 0.1 mL of candidate compound solutionappropriately diluted to obtain the desired concentration. A 1% controlvial, 0.1 mL of the 100-fold dilution of the inoculum described above,is inoculated into 12B vial without candidate compound. The 12B vialsare incubated at 37° C., and GI readings recorded daily, using a BACTEC460 TB instrument (Johnston Laboratories, Townsend, Md.), until thecontrol vial reaches a GI greater than 30. When the final readings inthe GI of the candidate containing vials are lower than those of the 1%control, the drug is considered to have inhibited more than 99% of thebacterial population, and this concentration will be defined as the MIC.

Table 1 gives MIC values for some of the compounds of the invention.TABLE 1 Structures and MIC values Com- MIC (μg/mL) pound No. Structure*MW MP (° C.) S. aureus S. pneumo. E. faecalis H. flu E. coli 1

971.06 226-230 0.008 0.00025 2 2 >8 2

1041.23 230-231 1 0.03 >8 >8 >8 3

929.027 206-216 0.03 0.0005 0.12 0.25 4 4

896.985 >300 0.03 0.00025 0.5 0.25 >8 5

925.039 >300 0.015 0.00025 0.5 2 8 6

1031.16 224-228 0.015 0.00025 0.25 1 4 7

883.002 240-243 0.12 0.004 4 4 >8 8

989.126 214-216 0.008 0.001 0.12 0.5 8 9

969.092 228-230 0.015 0.001 2 2 8 10

911.012 210-212 0.008 0.00012 1 2 >8 11

927.055 222-224 0.004 0.00012 0.25 1 4 12

925.039 >320 0.004 0.00025 0.12 0.25 8 13

999.117 184-188 0.008 0.00012 0.5 2 4 14

957.081 173-180 15

883.002 216-227 0.03 0.002 0.25 1 >8 16

868.975 208-229 0.015 0.001 0.12 0.25 4 17

927.055 >400 0.008 0.00012 1 1 8 18

885.018 214-216 0.004 0.00012 0.12 0.5 4 19

968.107 220-240 0.004 0.001 0.06 0.25 4 20

854.949 211-231 2 1 >8 >8 >8 21

969.048 >350 0.004 0.001 0.12 0.5 4 22

985.091 >300 0.015 0.00025 4 2 >8 23

1031.06 238-240 0.015 0.00012 4 2 >8 24

969.092 222-226 0.015 0.001 2 2 >8 25

977.092 256-257 0.008 0.001 2 2 >8 26

942.07 265-266 0.008 0.00012 2 1 >8 27

935.055 270-273 0.015 0.001 1 0.12 >8 28

984.107 260-263 0.03 0.002 1 0.25 >8 29

943.054 222-225 0.008 0.00012 2 1 >8 30

915 212-225 0.015 0.002 1 0.5 >8 31

926.071 250-252 0.03 0.0005 0.25 0.25 8 32

957.037 149-153 2 4 >8 >8 >8 33

899.001 278-279 0.008 0.00006 1 1 >8 34

970.08 234-236 0.03 0.001 2 0.5 8 35

1005.15 220-226 0.015 0.00025 4 0.5 8 36

927.055 228-230 0.004 0.00003 2 0.25 >8 37

928.043 321-322 0.03 0.004 2 0.25 >8 38

989.026 245-246 0.12 0.015 8 0.25 >8 39

856.964 265-266 0.03 0.001 2 0.25 >8 40

997.102 >300 0.06 0.008 1 0.5 >8 41

1001.09 175-183 0.25 0.0005 >8 2 >8 42

955.065 >300 0.008 0.00012 2 0.5 8 43

899.001 232-236 0.03 0.002 2 0.5 >8 44

941.038 232-234 0.06 0.008 2 0.25 >8 45

973.104 150-155 0.015 0.001 4 0.5 8 46

925.039 148-150 0.004 0.00006 2 0.5 >8 47

967.076 148-149 0.008 0.002 1 0.25 >8 48

999.117 218-228 0.008 0.00025 2 1 >8 49

1099.2 >300 0.06 0.0005 8 2 >8 50

1025.16 >300 0.03 0.00012 4 0.5 >8 51

985.091 175-185 0.06 0.002 8 0.25 8 52

957.081 230-232 0.015 0.00025 4 0.12 >8 53

925.039 >370 0.06 0.008 4 2 >8 54

883.002 >330 0.008 0.0005 2 0.25 >8 55

955.065 214-218 0.008 0.00025 1 0.5 8 56

913.028 205-213 0.004 0.001 0.12 0.25 8 57

960.084 220-222 0.008 0.0005 2 0.5 8 58

939.066 >300 0.03 0.004 1 0.25 >8 59

897.029 >300 0.004 0.001 0.06 0.12 4 60

943.054 110-195 0.008 0.0005 0.5 0.06 >8 61

928.039 205-212 0.004 0.00012 1 0.25 >8 62

918.047 228-230 0.015 0.008 8 0.5 >8 63

959.053 210-214 0.008 0.002 1 0.25 8 64

913.096 >250 0.004 0.00012 0.12 0.25 4 65

871.059 >300 0.002 0.00025 0.12 0.12 2 66

886.002 115-122 0.004 0.0005 0.12 0.12 4 67

963.109 0.008 0.001 0.5 0.12 >8 68

967.187 >250 0.06 0.0005 2 0.5 >8 69

927.123 >250 0.015 0.00025 0.5 0.5 4 70

925.151 >250 0.03 0.001 1 1 >8 71

941.082 216-220 0.008 0.00012 0.25 0.5 4 72

955.065 210-214 0.008 0.00025 2 0.5 >8 73

899.045 215-218 0.002 0.00025 0.03 0.12 2 74

913.028 220-221 0.03 0.001 0.5 0.12 >8 75

913.028 0.002 0.00012 0.12 0.25 4 76

925.107 >250 0.008 0.00025 0.5 0.25 4 77

883.07 >250 0.002 0.00025 0.25 0.12 2 78

875.99 0.002 0.00012 0.12 0.12 4 79

833.95*A, B, C, D, E, and F represent the following moieties:

Other Embodiments

All publications and patents cited in this specification are hereinincorporated by reference as if each individual publication or patentwere specifically and individually indicated to be incorporated byreference. Although the foregoing invention has been described in somedetail by way of illustration and example for purposes of clarity ofunderstanding, it will be readily apparent to those of ordinary skill inthe art in light of the teachings of this invention that certain changesand modifications may be made thereto without departing from the spiritor scope of the appended claims.

While the invention has been described in connection with specificembodiments, it will be understood that it is capable of furthermodifications. Therefore, this application is intended to cover anyvariations, uses, or adaptations of the invention that follow, ingeneral, the principles of the invention, including departures from thepresent disclosure that come within known or customary practice withinthe art.

Other embodiments are within the claims.

1. A compound having the Formula (I):

or a pharmaceutically acceptable salt thereof, wherein A is H, OH,O—(C₁-C₆ alkyl), or O—(C₁-C₄ alkaryl); W is O, S, or NR¹, wherein R¹ isH or C₁-C₆ alkyl; X is H or COR², wherein R² is C₁-C₆ alkyl, which canbe substituted with from 1 to 5 hydroxyl groups, or O—(C₃-C₇ alkyl),which can be substituted with from 1 to 4 hydroxyl groups, wherein eachalkyl carbon is bound to no more than one oxygen; Y is H, OR³, or Hal; Zis H, OR³, or Hal; R³ is C₁-C₆ alkyl; and R⁴ has the formula:

wherein, when each of m and n is 1, each of R⁵ and R⁶ is H, or R⁵ and R⁶together are ═O; R⁷ and R¹⁰ together form a single bond or a C₁-C₃linkage, which optionally contains a non-vicinal O, S, or N(R²³), R⁷ andR¹² together form a single bond or a C₁-C₂ linkage, which optionallycontains a non-vicinal O, S, or N(R²³), or R⁷ and R¹⁴ together form asingle bond or a C₁ linkage, wherein R²³ is H, C₁-C₆ alkyl, COR²⁴,CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴,wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, orC₁-C₄ alkheteroaryl; R⁸ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R⁸ andR¹² together form a single bond, or R⁸ and R⁹ together are ═N—O R¹⁸,wherein R¹⁸ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl; R⁹ is H, C₁-C₆ alkyl,or C₁-C₄ alkaryl, or R⁹ and R⁸ together are ═N—OR¹⁸; R¹⁰ is H, C₁-C₆alkyl, or C₁-C₄ alkaryl, or R¹⁰ and R⁷ together form a single bond or aC₁-C₃ linkage, which optionally contains a non-vicinal O, S, or N(R²³),or R¹⁰ and R¹⁶ together form a C₁-C₂ alkyl linkage, which optionallycontains a non-vicinal O, S, or N(R²³), or R¹⁰ and R¹⁷ together form aC₁-C₃ alkyl linkage, which optionally contains a non-vicinal O, S, orN(R²³), or R¹⁰ and R¹¹ together are ═O, wherein R²³ is H, C₁-C₆ alkyl,COR , CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, orSO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl,heteroaryl, or C₁-C₄ alkheteroaryl; R¹¹ is H, or R¹¹ and R¹⁰ togetherare ═O; R¹² is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R¹² and R⁷togetherform a single bond or a C₁-C₂ linkage, which optionally contains anon-vicinal O, S, or N(R²³), or R¹² and R⁸ together form a single bond,or R¹² and R¹⁶ together form a C₂-C₄ alkyl linkage, which optionallycontains a non-vicinal O, S, or N(R²³), wherein R²³ is H, C₁-C₆ alkyl,COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R , orSO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl,heteroaryl, or C₁-C₄ alkheteroaryl; each of R¹³ and R¹⁵ is H, C₁-C₆alkyl, or C₁-C₄ alkaryl; R¹⁴ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R¹⁴and R⁷ together form a single bond or a C₁ linkage; R¹⁶ is H, C₁-C₆alkyl, C₁-C₆ alkoxy, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl, or C₁-C₄alkheteroaryl, or R¹⁶ and R¹² together form a C₂-C₄ alkyl linkage, whichoptionally contains a non-vicinal O, S, or N(R²³), or R¹⁶ and R¹⁰together form a C₁-C₂ alkyl linkage, which optionally contains anon-vicinal O, S, or N(R²³), wherein R²³ is H, C₁-C₆ alkyl, COR²⁴,CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴,wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, orC₁-C₄ alkheteroaryl; and R¹⁷ is H, C₁-C₆ alkyl, COR¹⁹, CO₂R¹⁹, orCONHR¹⁹, CSR¹⁹, COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or SO₂NHR¹⁹, whereinR¹⁹ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄alkheteroaryl, or R¹⁷ and R¹⁰ together form a C₁-C₃ alkyl linkage, whichoptionally contains a non-vicinal O, S, or N(R²³), wherein R²³ is H,C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴,SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl; or when m is 0 and n is 1,R⁷ and R¹⁰ together form a single bond or a C₁-C₄ linkage, whichoptionally contains a non-vicinal O, S, or N(R²³), R⁷ and R¹² togetherform a single bond or a C₁-C₃ linkage, which optionally contains anon-vicinal O, S, or N(R²³), or R⁷ and R¹⁴ together form a single bondor a C₁-C₂ linkage, which optionally contains a non-vicinal O, S, orN(R²³), wherein R²³ is H, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴,COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl;each of R⁸, R⁹, and R¹¹ is H; R¹⁵ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl;R¹⁰ is H or R¹⁰ and R⁷ together form a single bond or a C₁-C₄ linkage,which optionally contains a non-vicinal O, S, or N(R²³), wherein R²³ isH, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴,CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl,C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl; R¹² is H, C₁-C₆alkyl, or C₁-C₄ alkaryl, or R¹² and R⁷ together form a single bond or aC₁-C₃ linkage, which optionally contains a non-vicinal O, S, or N(R²³),R¹² and R¹³ together form a —CH₂CH₂— linkage, or R¹² and R¹⁶ togetherform a C₂-C₄ alkyl linkage, which optionally contains a non-vicinal O,S, or N(R²³), wherein R²³ is H, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴,CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ isC₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄alkheteroaryl; R¹³ is H, C₁-C₆ alkyl, C₁-C₄ alkaryl, or R¹³ and R¹²together form a —CH₂CH₂— linkage; R¹⁴ is H, C₁-C₆ alkyl, or C₁-C₄alkaryl, or R¹⁴ and R⁷ together form a single bond or a C₁-C₂ linkage,which optionally contains a non-vicinal O, S, or N(R²³), wherein R²³ isH, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴,CSNHR²⁴, SO₂R²⁴, or SO₂NHR , wherein R is C₁-C₆ alkyl, C₆-C₁₂ aryl,C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl; R¹⁶ is H, C₁-C₆alkyl, C₁-C₆ alkoxy, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl, or C₁-C₄alkheteroaryl, or R¹⁶ and R¹² together form a C₂-C₄ alkyl linkage, whichoptionally contains a non-vicinal O, S, or N(R²³), wherein R²³ is H,C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴,SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl; and R¹⁷ is H, C₁-C₆ alkyl,COR¹⁹, CO₂R¹⁹, or CONHR¹⁹, CSR¹⁹, COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, orSO₂NHR¹⁹, wherein R¹⁹ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl,heteroaryl, or C₁-C₄ alkheteroaryl; or A is OH; X is H; W, Y, and Z aredefined as above; and R⁴ is selected from the group consisting of:

wherein R²¹ is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl,or C₁-C₄ alkheteroaryl, R²⁰ is H, C₁-C₆ alkyl, COR¹⁹, CO₂ ¹⁹, orCONHR¹⁹, CSR¹⁹, COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or SO₂NHR¹⁹, whereinR¹⁹ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄alkheteroaryl; or A is OH; X is COCH₃; W, Y, and Z are defined as above;and R⁴ is selected from the group consisting of:

wherein R²¹ is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl,or C₁-C₄ alkheteroaryl, R²⁰ is H, C₁-C₆ alkyl, COR¹⁹, CO₂R¹⁹, orCONHR¹⁹, CSR¹⁹, COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or SO₂NHR¹⁹, whereinR¹⁹ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄alkheteroaryl; or A is H or OH; X is H or COCH₃; W, Y, and Z are definedas above; and R⁴ is

with the proviso that one or both of Y and Z are Hal; or A is H or OH; Xis H or COCH₃; W, Y, and Z are defined as above; and R⁴ is

wherein R²² is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl,C₁-C₄ alkheteroaryl, COR²⁴, CO₂R²⁴, CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴,CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl,C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl, and r is 1-2; or A isH or OH; X is H or COCH₃; W, Y, and Z are defined as above; and R⁴ is

wherein R²¹ is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl,or C₁-C₄ alkheteroaryl; or A is H or OH; X is H or COCH₃; W, Y, and Zare defined as above; and R⁴ is

wherein ═E is ═O or (H,H), R²² is H, C₁-C₆ alkyl, C₆-C₁₂ aryl,heteroaryl, C₁-C₄ alkaryl, C₁-C₄ alkheteroaryl, COR²⁴, CO₂R²⁴, CONHR²⁴,CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴ or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl, ris 1-2, and s is 0-1; or A is H or OH; X is H or COCH₃; W, Y, and Z aredefined as above; and R⁴ is


2. The compound of claim 1, wherein, R⁴ has the formula:

wherein, when each of m and n is 1, each of R⁵ and R⁶ is H, or R⁵ and R⁶together are ═O; R⁷ and R¹⁰ together form a single bond or a C₁-C₃linkage, which optionally contains a non-vicinal O, S, or N(R²³), R⁷ andR¹² together form a single bond or a C₁-C₂ linkage, which optionallycontains a non-vicinal O, S, or N(R²³), or R⁷ and R¹⁴ together form asingle bond or a C₁ linkage, wherein R²³ is H, C₁-C₆ alkyl, COR²⁴,CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴ or SO₂NHR²⁴,wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, orC₁-C₄ alkheteroaryl; R⁸ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R⁸ andR¹² together form a single bond, or R⁸ and R⁹ together are ═N—O R¹⁸,wherein R¹⁸ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl; R⁹ is H, C₁-C₆ alkyl,or C₁-C₄ alkaryl, or R⁹ and R⁸ together are ═N—OR¹⁸; R¹⁰ is H, C₁-C₆alkyl, or C₁-C₄ alkaryl, or R¹⁰ and R⁷ together form a single bond or aC₁-C₃ linkage, which optionally contains a non-vicinal O, S, or N(R²³),or R¹⁰ and R¹⁶ together form a C₁-C₂ alkyl linkage, which optionallycontains a non-vicinal O, S, or N(R²³), or R¹⁰ and R¹⁷ together form aC₁-C₃ alkyl linkage, which optionally contains a non-vicinal O, S, orN(R²³), or R¹⁰ and R¹¹ together are ═O, wherein R²³ is H, C₁-C₆ alkyl,COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, orSO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl,heteroaryl, or C₁-C₄ alkheteroaryl; R¹¹ is H, or R¹¹ and R¹⁰ togetherare ═O; R¹² is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R¹² and R⁷ togetherform a single bond or a C₁-C₂ linkage, which optionally contains anon-vicinal O, S, or N(R²³), or R¹² and R⁸ together form a single bond,or R¹² and R¹⁶ together form a C₂-C₄ alkyl linkage, which optionallycontains a non-vicinal O, S, or N(R²³), wherein R²³ is H, C₁-C₆ alkyl,COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR₂₄, SO₂R²⁴, orSO₂NHR²⁴, wherein R₂₄ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl,heteroaryl, or C₁-C₄ alkheteroaryl; each of R¹³ and R¹⁵ is H, C₁-C₆alkyl, or C₁-C₄ alkaryl; R¹⁴ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R¹⁴and R⁷ together form a single bond or a C₁ linkage; R¹⁶ is H, C₁-C₆alkyl, C₁-C₆ alkoxy, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl, or C_(l)-C₄alkheteroaryl, or R¹⁶ and R¹² together form a C₂-C₄ alkyl linkage, whichoptionally contains a non-vicinal O, S, or N(R²³), or R¹⁶ and R¹⁰together form a C₁-C₂ alkyl linkage, which optionally contains anon-vicinal O, S, or N(R²³), wherein R²³ is H, C₁-C₆ alkyl, COR²⁴,CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴,wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, orC₁-C₄ alkheteroaryl; and R¹⁷ is H, C₁-C₆ alkyl, COR¹⁹, CO₂R¹⁹, orCONHR¹⁹, wherein R¹⁹ is C₁-C₆ alkyl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄alkheteroaryl, or R¹⁷ and R¹⁰ together form a C₁-C₃ alkyl linkage, whichoptionally contains a non-vicinal O, S, or N(R²³), wherein R²³ is H,C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴,SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl; or when m is 0 and n is 1,R⁷ and R¹⁰ together form a single bond or a C₁-C₄ linkage, whichoptionally contains a non-vicinal O, S, or N(R²³), R⁷ and R¹² togetherform a single bond or a C₁-C₃ linkage, which optionally contains anon-vicinal O, S, or N(R²³), or R⁷ and R¹⁴ together form a single bondor a C₁-C₂ linkage, which optionally contains a non-vicinal O, S, orN(R²³), wherein R²³ is H, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴,COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl;each of R⁸, R⁹, and R¹¹ is H; R¹⁵ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl;R¹⁰ is H or R¹⁰ and R⁷ together form a single bond or a C₁-C₄ linkage,which optionally contains a non-vicinal O, S, or N(R²³), wherein R²³ isH, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴,CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl,C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl; R¹² is H, C₁-C₆alkyl, or C₁-C₄ alkaryl, or R¹² and R⁷ together form a single bond or aC₁-C₃ linkage, which optionally contains a non-vicinal O, S, or N(R²³),R¹² and R¹³ together form a —CH₂CH₂— linkage, or R¹² and R¹⁶ togetherform a C₂-C₄ alkyl linkage, which optionally contains a non-vicinal O,S, or N(R²³), wherein R²³ is H, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴,CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ isC₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄alkheteroaryl; R¹³ is H, C₁-C₆ alkyl, C₁-C₄ alkaryl, or R¹³ and R¹²together form a —CH₂CH₂— linkage; R¹⁴ is H, C₁-C₆ alkyl, or C₁-C₄alkaryl, or R¹⁴ and R⁷ together form a single bond or a C₁-C₂ linkage,which optionally contains a non-vicinal O, S, or N(R²³), wherein R²³ isH, C₁-C₆ alkyl, COR²⁴, CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴,CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl,C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl; R¹⁶ is H, C₁-C₆alkyl, C₁-C₆ alkoxy, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl, or C₁-C₄alkheteroaryl, or R¹⁶ and R¹² together form a C₂-C₄ alkyl linkage, whichoptionally contains a non-vicinal O, S, or N(R²³), wherein R²³ is H,C₁-C₆ alkyl, COR²⁴, CO₂R, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴,SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl; and R¹⁷ is H, C₁-C₆ alkyl,COR¹⁹, CO₂R¹⁹, or CONHR¹⁹, CSR¹⁹, COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, orSO₂NHR¹⁹, wherein R¹⁹ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl,heteroaryl, or C₁-C₄ alkheteroaryl.
 3. The compound of claim 1, whereinA is OH, X is H, and R⁴ is selected from the group consisting of:


4. The compound of claim 1, wherein A is OH, X is COCH₃, and R⁴ isselected from the group consisting of:


5. The compound of claim 1, wherein A is H or OH, X is H or COCH₃, andR⁴ is

wherein one or both of Y and Z is F.
 6. The compound of claim 1, whereinW is O.
 7. The compound of claim 1, wherein W is S.
 8. The compound ofclaim 1, wherein W is NR¹, wherein R¹ is H or C₁-C₆ alkyl.
 9. Thecompound of claim 2, wherein A is OH.
 10. The compound of claim 1,wherein X is COR², wherein R² is O(C₃-C₇ alkyl), which can besubstituted with from 1 to 4 hydroxyl groups, wherein each alkyl carbonis bound to no more than one oxygen.
 11. The compound of claim 1,wherein W is O; Y is H; Z is H; A is OH; X is H or COCH₃; and R⁴ is

wherein each of R⁵ and R⁶ is H, or R⁵ and R⁶ together are ═O; each ofR⁸, R⁹, R¹², R¹³ and R¹⁵ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl; each ofR¹⁰ and R¹¹ is H, C₁-C₆ alkyl, or C₁-C₄ alkaryl, or R¹⁰ and R¹⁰ togetherare ═O; and R¹⁷ is H, C_(1-C) ₆ alkyl, COR¹⁹, CO₂R¹⁹, or CONHR¹⁹, CSR¹⁹,COSR¹⁹, CSOR¹⁹, CSNHR¹⁹, SO₂R¹⁹, or SO₂NHR¹⁹, wherein R¹⁹ is C₁-C₆alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl.12. The compound of claim 11, wherein R⁴ is


13. The compound of claim 11, wherein R⁴ is


14. The compound of claim 1, wherein W is O; Y is H; Z is H; X is H orCOCH₃; A is H or OH; and R⁴ is selected from the group consisting of:


15. The compound of claim 1, wherein W is O; Y is H; Z is H; X is COCH₃;A is OH; and R⁴ is:


16. The compound of claim 1, wherein W is O; Y is H; Z is H; X is COCH₃;A is OH; and R⁴ is:

wherein each of R¹⁷ and R²³ is, independently H, C₁-C₆ alkyl, COR²⁴,CO₂R²⁴, or CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴,wherein R₂₄ is C₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, orC₁-C₄ alkheteroaryl.
 17. The compound of claim 1, wherein W is O; Y isH; Z is H; A is H or OH; X is H or COCH₃; and R⁴ is

wherein R²¹ is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl,or C₁-C₄ alkheteroaryl.
 18. The compound of claim 1, wherein W is O; Yis H; Z is H; A is H or OH; X is H or COCH₃; and R⁴ is

wherein R²² is H, C₁-C₆ alkyl, C₆-C₁₂ aryl, heteroaryl, C₁-C₄ alkaryl,C₁-C₄ alkheteroaryl, COR²⁴, CO₂R²⁴, CONHR²⁴, CSR²⁴, COSR²⁴, CSOR²⁴,CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ is C₁-C₆ alkyl, C₆-C₁₂ aryl,C₁-C₄ alkaryl, heteroaryl, or C₁-C₄ alkheteroaryl, and r is 1 or
 2. 19.The compound of claim 1, wherein W is O; Y is H; Z is H; A is H or OH; Xis H or COCH₃; and R⁴ is

wherein ═E is ═O or (H,H), R²² is H, C₁-C₆ alkyl, C₆-C₁₂ aryl,heteroaryl, C₁-C₄ alkaryl, C₁-C₄ alkheteroaryl, COR²⁴, CO₂R²⁴, CONHR²⁴,CSR²⁴, COSR²⁴, CSOR²⁴, CSNHR²⁴, SO₂R²⁴, or SO₂NHR²⁴, wherein R²⁴ isC₁-C₆ alkyl, C₆-C₁₂ aryl, C₁-C₄ alkaryl, heteroaryl, or C₁-C₄alkheteroaryl, r is 1-2, and s is 0-1.
 20. The compound of claim 1,wherein W is O; Y is H; Z is H; A is H or OH; X is H or COCH₃; and R⁴ is


21. The compound of claim 1, wherein said compound of formula (I) isselected from:3′-hydroxy-5′-(4-ethylcarbamyl-1-piperidinyl)benzoxazinorifamycin,3′-hydroxy-5′-[6-(2-trimethylsilyl)ethylcarbamyl-(1R,5S)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-(4-ethylcarbamyl-1-piperidinyl)benzoxazinorifamycin,3′-hydroxy-5′-[6-amino-(1R,5S)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,3′-hydroxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,3′-hydroxy-5′-(1-piperidinyl-4-(N-phenyl)propanamide)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-(1-piperidinyl-4-(N-phenyl)propanamide)benzoxazinorifamycin,3′-hydroxy-5′-(4-morpholinyl-1-piperidinyl)benzoxazinorifamycin,3′-hydroxy-5′-(3,8-diazabicyclo[3.2.1]octan-3-yl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-(4-morpholinyl-1-piperidinyl)benzoxazinorifamycin,3′-hydroxy-5′-[(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,3′-hydroxy-5′-(4-(2-methylpropyl)carbamyl-1-piperidinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-(4-(2-methylpropyl)carbamyl-1-piperidinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-(3,8-diazabicyclo[3.2.1]octan-3-yl)benzoxazinorifamycin,3′-hydroxy-5′-(4-N,N-dimethylamino-1-piperidinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-(4-N,Ndimethylamino-1-piperidinyl)benzoxazinorifamycin,5′-(4-ethylcarbamyl-1-piperidinyl)-N′-methylbenzodiazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[6-amino-(1R,5S)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,3′-hydroxy-5′-[6-ethylcarbamyl-(1R,5S)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,3′-hydroxy-5′-[4-isopropylcarbamyl-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-trifluoromethylsulfonyl-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-butanamide-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-methylsulfonyl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-propyluryl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-methylsulfonyl-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-propyluryl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-isopropylcarbamyl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-methylcarbamyl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-5′-(4-ethylcarbamyl-1-piperidinyl)-N′-methylbenzdiazinorifamycin,3′-hydroxy-5′-[4-methylcarbamyl-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-amino-i-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-ethyluryl-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-propylsulfonyl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-butanamide-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-ethyluryl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-trifluoromethysulfonyl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-amino-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[1-ethylcarbamyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,3′-hydroxy-5′-[1-ethylcarbamyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,3′-hydroxy-5′-[4-methoxyethylcarbamyl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[1-ethylcarbamyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[1-ethylcarbamyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-acetamide-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-acetyl-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-S-methylthiocarbamyl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[1-acetyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[1-acetyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,3′-hydroxy-5′-[1-acetyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,3′-hydroxy-5′-[1-acetyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,3′-hydroxy-5′-[4-(2,2-dimethylethyl)carbamyl-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-(4-(S-methylthiocarbamyl)-1-piperidinylcarbonyl)amino-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-(4-methylpiperazinylcarbonyl)amino-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-ethylcarbamylmethyl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-(2,2-dimethylethyl)carbamyl-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[6-N,N-dimethylamino-(1R,5S)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,3′-hydroxy-5,′-[6-N,N-dimethylamino-(1R,5S)-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,3′-hydroxy-5′-[4-acetylaminomethyl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-acetylaminomethyl-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-phenyl-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[1-methyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,3′-hydroxy-5′-[1-methyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[1-methyl-(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[1-methyl-(4aR,7aR)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-ethylcarbamylmethyl-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-(2-hydroxyethyl)-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-phenyl-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-methoxyethylcarbamyl-1-piperidinylbenzoxazinorifamycin,5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin,5′-[(3S,5R)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin,25-O-deacetyl-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin,25-O-deacetyl-5′-[(3S,5R)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-(2-hydroxyethyl)-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-propylsulfonyl-1-piperidinyl]benzoxazinorifamycin,5′-[(2S,5R)-4-(cyclopropylmethyl)-2,5-dimethylpiperazinyl]benzthiazinorifamycin,5′-[(2R,5S)-4-(cyclopropylmethyl)-2,5-dimethylpiperazinyl]benzthiazinorifamycin,5′-[4-N,N-dimethylamino-1-piperidinyl]benzthiazinorifamycin,25-O-deacetyl-5′-[(2S,5R)-4-(cyclopropylmethyl)-2,5-dimethylpiperazinyl]benzthiazinorifamycin,25-O-deacetyl-5′-[(2R,5S)-4-(cyclopropylmethyl)-2,5-dimethylpiperazinyl]benzthiazinorifamycin,3′-hydroxy-5′-[4-methyl-4-N,N-dimethylamino-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[4-methyl-4-acetylamino-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-methyl-4-N,N-dimethylamino-1-piperidinyl]benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-[4-methyl-4-acetylamino-1-piperidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[(3R)-N,N-dimethylamino-1-pyrrolidinyl]benzoxazinorifamycin,3′-hydroxy-5′-[(3S)-N,N-dimethylamino-1-pyrrolidinyl]benzoxazinorifamycin,5′-[(8aS)octahydropyrrolo[1,2-a]pyrazin-2-yl]benzthiazinorifamycin,5′-[(8aR)octahydropyrrolo[1,2-a]pyrazin-2-yl]benzthiazinorifamycin,25-O-deacetyl-5′-[(8aS)octahydropyrrolo[1,2-a]pyrazin-2-yl]benzthiazinorifamycin,25-O-deacetyl-5′-[(8aR)octahydropyrrolo[1,2-a]pyrazin-2-yl]benzthiazinorifamycin,3′-hydroxy-5′-[3-hydroxy-1-azetidinyl]benzoxazinorifamycin, and25-O-deacetyl-3′-hydroxy-5′-[3-hydroxy-1-azetidinyl]benzoxazinorifamycin.22. The compound of claim 1, wherein said compound of formula (I) isselected from:4′-fluoro-5′-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin,4′-fluoro-5′-(1-piperazinyl)benzoxazinorifamycin,4′-fluoro-5′-(3-methyl-1-piperazinyl)benzoxazinorifamycin,4′-methoxy-6′-fluoro-5′-(3-methyl-1-piperazinyl)benzoxazinorifamycin,4′,6′-difluoro-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzoxazinorifamycin,4′-fluoro-6′-methoxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,4′-fluoro-5′-[6-amino-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(3-methyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-4′-methoxy-6′-fluoro-5′-(3-methyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-4′,6′-difluoro-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzoxazinorifamycin,25-O-deacetyl-4′-fluoro-6′-methoxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-4′-fluoro-5′-[6-amino-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-5′-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-25-(2 ″,3″-dihydroxypropylcarbonoxy)-4′-fluoro-5′-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-fluoro-5′-(1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-fluoro-5′-(3-methyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-methoxy-6′-fluoro-5′-(3-methyl-1-piperazinyl)benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′,6′-difluoro-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-fluoro-6′-methoxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzoxazinorifamycin,25-O-deacetyl-25-(2″,3″-dihydroxypropylcarbonoxy)-4′-fluoro-5′-[6-amino-3-azabicyclo[3.1.0]hex-3-yl]benzoxazinorifamycin,4′-fluoro-5′-(4-isobutyl-1-piperazinyl)benzthiazinorifamycin,4′-fluoro-5′-(1-piperazinyl)benzthiazinorifamycin,4′-fluoro-5′-(3-methyl-1-piperazinyl)benzthiazinorifamycin,4′-methoxy-6′-fluoro-5′-(3-methyl-1-piperazinyl)benzthiazinorifamycin,4′,6′-difluoro-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin,4′-fluoro-6′-methoxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzthiazinorifamycin,4′-fluoro-5′-[6-amino-3-azabicyclo[3.1.0]hex-3-yl]benzthiazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(4-isobutyl-1-piperazinyl)benzthiazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(1-piperazinyl)benzthiazinorifamycin,25-O-deacetyl-4′-fluoro-5′-(3-methyl-1-piperazinyl)benzthiazinorifamycin,25-O-deacetyl-4′-methoxy-6′-fluoro-5′-(3-methyl-1-piperazinyl)benzthiazinorifamycin,25-O-deacetyl-4′,6′-difluoro-5′-[(3R,5S)-3,5-dimethyl-1-piperazinyl]benzthiazinorifamycin,25-O-deacetyl-4′-fluoro-6′-methoxy-5′-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]benzthiazinorifamycin,25-O-deacetyl-4′-fluoro-5′-[6-amino-3-azabicyclo[3.1.0]hex-3-yl]benzthiazinorifamycin,3′-hydroxy-5′-((3R,5S)-3,5-dimethylpiperazinyl)benzoxazinorifamycin,3′-hydroxy-5′-((3R,5S)-3 ,5-diethylpiperazinyl)benzoxazinorifamycin,3′-hydroxy-5′-((3R,5S)-3-ethyl-5-methylpiperazinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-((3R,5S)-3,5-dimethylpiperazinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-((3R,5S)-3-ethyl-5-methylpiperazinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-((3R,5S)-3,5-diethylpiperazinyl)benzoxazinorifamycin,3′-hydroxy-5′-((4aR,7aR)octahydro-1H-pyrrolyl[3,4-b]pyridine)benzoxazinorifamycin,3′-hydroxy-5′-((4aS,7aS)octahydro-1H-pyrrolyl[3,4-b]pyridine)benzoxazinorifamycin,3′-hydroxy-5′-((8aR)-octahydropyrrolyl[1,2-a]pyrazinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-((8aR)-octahydropyrrolyl[1,2-a]pyrazinyl)benzoxazinorifamycin,3′-hydroxy-5′-((8aS)-octahydropyrrolyl[1,2-a]pyrazinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-((8aS)-octahydropyrrolyl[1,2-a]pyrazinyl)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-(4-methylpiperazinyl)benzoxazinorifamycin,3′-hydroxy-5′-(ethyl piperidinyl-4-ylcarbamate)benzoxazinorifamycin,25-O-deacetyl-3′-hydroxy-5′-(ethylpiperidinyl-4-ylcarbamate)benzoxazinorifamycin,3′-hydroxy-5′-((3Z)-4-(aminomethyl)pyrrolidinyl-3-one O-methyloxime)benzoxazinorifamycin, 3′-hydroxy-5′-(5-azaspiro[2.4]heptan-7-amino-5-yl)benzoxazinorifamycin, and 3′-hydroxy-5′-(5-aminopyrrolidinyl)benzoxazinorifamycin.
 23. A pharmaceutical composition comprising apharmaceutically acceptable carrier or diluent and a compound of formula(I).
 24. The composition of claim 23, wherein said composition is (i) acompound of formula I in an amount between 0.001% and 5% weight/volume(w/v) and (ii) a pharmaceutically acceptable carrier suitable fortopical administration to a patient.
 25. A method of treating, orpreventing a microbial infection in an animal, said method comprisingadministering to said animal a pharmaceutical composition of claim 23 inan amount sufficient to treat or prevent said microbial infection. 26.The method of claim 25, wherein said pharmaceutical composition isadministered orally, topically, intravenously, intramuscularly, orsubcutaneously.
 27. The method of claim 25, wherein said infection is aprotozoan, bacterial, viral, or fungal infection.
 28. The method ofclaim 25, wherein said animal is a human patient.
 29. The method ofclaim 28, wherein said microbial infection is Clostridium difficile. 30.The method of claim 28, wherein said microbial infection is Chlamydiatrachomatis or N. gonorrhoeae.
 31. The method of claim 28, wherein saidmicrobial infection is H. pylori.